Integrated Biological Protection Cell Sorter

Abstract

An integrated bioprotective cell sorter is disclosed, which isolates the potentially contaminating parts of the cell sorter and includes two protection systems. The main cabinet protection system contains the input sample. The aerosol management protection area includes a nozzle chamber having nozzles, a sorting chamber having a sorting plate, and a collection medium for collecting the droplet stream from the nozzles. The main cabinet is maintained at a first low pressure, and clean air circulates under positive pressure. The aerosol management protection area is maintained at a second low pressure lower than the first pressure, so that contaminants do not leak from the aerosol management protection area into the main cabinet protection area. A sliding sash is located above the entrance and exit in the main cabinet and can be moved to access different parts of the main cabinet without changing the substantially constant first low pressure in the main cabinet.

Description

Background Art

[0001] Cell sorters and flow cytometers have become important laboratory tools. A cell sorter can identify certain types of biological cells and separate these cells from other cells. Commercial uses of cell sorters have also been realized in multiple industries. There are many other uses for cell sorters, such as identifying and separating various types of cells for laboratory applications. Therefore, cell sorters have many different and diverse uses and applications. Summary of the Invention

[0002] Accordingly, one embodiment of the present invention may include an integrated biohazard cell sorter flow cytometer, comprising: a main cabinet of the integrated biohazard cell sorter, which is not airtight; an input sample area disposed within the main cabinet; a movable partition located within the main cabinet and moving within the entrance and exit of the main cabinet; when the movable partition moves within the entrance and exit, the movable partition covers a constant area of the entrance and exit, such that when the movable partition moves within the entrance and exit, a constant area of the entrance and exit is not covered by the movable partition and is open; a first fan that sucks air from the main cabinet to create a first low pressure within the main cabinet, and the first low pressure is substantially constant when the movable partition moves within the entrance and exit, which restricts contaminated air within the main cabinet from escaping from the main cabinet; an aerosol management protection area, which is not airtight and is disposed within the main cabinet; the aerosol management protection area has an opening that connects to the main cabinet such that the aerosol management protection area is disposed within the first low pressure and is subject to the first low pressure; the aerosol management protection area has a nozzle that generates a stream of droplets containing sample cells, a sorting plate that divides the stream of droplets into deflected streams, and a collection medium for collecting the deflected streams; a second fan that sucks air from the aerosol management protection area to create a second low pressure within the aerosol management protection area that is lower than the first low pressure, such that air from the main cabinet flows from the main cabinet to the aerosol management protection area and restricts contaminated air from flowing from the aerosol management protection area into the main cabinet; a light excitation device located outside the main cabinet and the aerosol management protection area to allow access to the light excitation device without entering the main cabinet or the aerosol management protection area.

[0003] Accordingly, another embodiment of the present invention may include a method of intercepting cells in an integrated biocontainment cell sorter, which includes: providing a main cabinet protection area containing an input area for sample cells to be sorted; generating a first low pressure in the main cabinet using a first fan that draws air from the main cabinet and from outside the cabinet; generating a second low pressure in an aerosol management protection area provided in the main cabinet using a second fan that draws air from the main cabinet and the aerosol management protection area through an opening in the aerosol management protection area to generate the second low pressure lower than the first low pressure in the aerosol management protection area; enclosing the input cell sample in the main cabinet that is not airtight sealed; enclosing the nozzle, sorting plate, collection medium, and any droplet stream generated by the nozzle in the aerosol management protection area that is not airtight sealed; and disposing a light excitation device outside the main cabinet and the aerosol management protection area to facilitate adjustment and maintenance of the light excitation device. BRIEF DESCRIPTION OF THE DRAWINGS

[0004] Figure 1 is a schematic side cross-sectional view showing the various parts of an embodiment of an integrated biocontainment cell sorter.

[0005] Figure 2 is Figure 1 a schematic diagram of the aerosol management system (AMS) thereof, the system including a nozzle chamber and a sorting chamber and the various other parts of an embodiment of an integrated biocontainment cell sorter.

[0006] Figure 3 is a schematic side cross-sectional view showing Figure 2 the nozzle chamber and the sorting chamber in more detail.

[0007] Figure 4 is a detailed front view of an embodiment of an integrated biocontainment cell sorter, which shows Figure 2 and 3 the nozzle chamber and the sorting chamber.

[0008] Figure 5 is Figures 1 to 4 a perspective view of a specific implementation of the integrated biocontainment cell sorter shown in DETAILED DESCRIPTION

[0009] Figure 1Schematic diagram of an embodiment of an integrated biological protection system 100. The system consists of a main cabinet protection system 101 and an aerosol management system chamber 140, an AMS intake pipe 138, an AMS HEPA filter 141, an AMS fan 144, and an AMS exhaust pipe 147, which are collectively referred to as the aerosol management system (AMS) 149. The aerosol management system (AMS) 149 is contained in or connected to the main cabinet protection system 101. Neither the main cabinet protection system 101 nor the aerosol management system (AMS) 149 is a sealed system. Instead, they rely on the air movement caused by the fan 122 in the main cabinet protection system 101 and the AMS fan 144 in the aerosol management system (AMS) 149 to generate a low pressure, so that harmful substances and toxic substances do not diffuse outside each of these protection systems. For example, since the fan 122 draws air from the working area 104 of the main cabinet protection system 101 and outside the main cabinet protection system 101, a first low pressure is generated in the working area 104, as shown by the air flow 116. Thus, a first low pressure is generated in the working area 104 where potentially toxic or hazardous substances may be present. As long as the fan 122 can inhale enough air through the working area 104, the hazardous substances will not escape from the working area 104 to the area outside the main cabinet protection system 101. The hazardous substances mainly exist in the form of aerosols containing sample cells. The sample cells are mixed with the sheath fluid and pass through the nozzle 146. Generally, the nozzle generates a nozzle flow 153 ( Figure 2 ) that breaks into a droplet flow 155 ( Figure 2 ). If the nozzle is blocked, then aerosols containing sample cells will be generated. In addition, if the nozzle flow 153 or the droplet flow 155 impacts a hard surface, then aerosols will be generated. The aerosols contain sample cells that should not be inhaled or ingested. For example, the sample cells can be cancer cells.

[0010] Similarly, as Figure 1As shown, the air inhaled by the fan 122 first passes through the HEPA filter 120 to remove any hazardous substances, such as sample cells. Thus, the air inhaled by the fan 122 is clean air and the fan is not contaminated. The fan 122 forces the clean air to pass through the recirculation duct 108 under positive pressure, which also remains clean. Even under positive pressure, the recirculation duct 108 and the recirculation plenum chamber do not have to be sealed because they contain clean air. A portion of the air from the fan 122 is exhausted from the exhaust port 110, denoted as exhaust 126. At the same time, some of the air from the fan 122 is recirculated, denoted as recirculated air 124. Thus, the recirculation duct 108 circulates clean air under positive pressure, and the clean air is recirculated under positive pressure into the recirculation plenum chamber 102, while the remaining air is exhausted from the exhaust port 110. The recirculated air in the recirculation duct 108 and the recirculation plenum chamber 102 is under positive pressure and may thus leak from these ducts into the outside air or other parts of the integrated biological protection system 100. Since the air under positive pressure is clean air, unlike many other protection systems, there is no contamination problem with this system. The recirculated air in the recirculation plenum chamber 102 passes through the air flow straightener 128. The air flow straightener 128 is a device with openings that cause the recirculated air to flow into the work area 104 as a substantially uniform laminar flow 130 with low turbulence. The low turbulence allows for the maintenance of a uniform downward air volume, which both prevents contaminants from the inside of the cabinet from escaping through the user access opening and prevents contaminants from outside the cabinet from depositing on the products inside the cabinet.

[0011] The aerosol management system chamber 140 has an opening connected to the main cabinet protection system 101, also as Figure 1As shown. The Aerosol Management System (AMS) 149 has a separate AMS HEPA filter 141 and a separate AMS fan 144. The AMS fan 144 sucks air from the AMS intake pipe 138 connected to the sorting chamber 131. Since the AMS chamber 140 is connected to the main cabinet protection system 101 through an opening, the AMS chamber 140 is already under the first low pressure maintained in the main cabinet protection system 101. The AMS fan 144 further reduces the pressure in the Aerosol Management System (AMS) 149 from the first low pressure in the working area 104 to a second low pressure lower than the first low pressure. When the nozzle chamber door 136 or the sorting chamber door 132 is opened, the second low pressure in the AMS chamber 140 will be equal to the first low pressure in the main cabinet protection system 101. Therefore, due to the opening of the nozzle chamber door 136 or the sorting chamber door 132, the pressures of the main cabinet protection system 101 and the AMS chamber 140 are equal, and air initially flows from the main cabinet protection system 101 to the AMS chamber 140, which prevents aerosols from escaping from the AMS chamber 140. However, once the pressure is balanced, the aerosols will migrate from the AMS chamber 140 to the main cabinet protection system 101. Therefore, before opening the nozzle chamber door 136 or the sorting chamber door 132, the nozzle 146 is closed and the AMS fan 144 runs at an accelerated speed for a period of time to discharge all aerosols from the AMS chamber 140. During operation, with the nozzle chamber door 136 and the sorting chamber door 132 closed, the second low-pressure air in the AMS chamber 140 sucks air from the working area 104 of the main cabinet protection system 101 at the AMS inlets 135, 137, and 160. In other words, the air at the first low pressure in the main cabinet protection system 101 in the working area 104 is sucked into the AMS chamber 140 because when the nozzle chamber door 136 and the sorting chamber door 132 are closed, the second low pressure in the AMS chamber 140 is lower than the first low pressure in the working area 104. Similarly, this is the result of the AMS fan 144 sucking air from the AMS chamber 140 through the AMS HEPA filter 141 via the AMS intake pipe 138 into the external environment. The clean air that has been filtered by the AMS HEPA filter 141 and sucked by the AMS fan 144 is discharged to the exhaust port 110 through the AMS exhaust pipe 147. Therefore, the Aerosol Management System (AMS) 149 is not only a protection system parallel to the main cabinet protection system 101, but the AMS chamber 140 is connected to the main cabinet protection system 101 through the intake port to generate a second low pressure, which makes it more difficult for hazardous substances located in the AMS chamber 140 and the working area 104 to escape from the integrated biological protection system 100.

[0012] As Figure 1As further shown, the aerosol management system chamber 140 is carefully constructed to enclose the portion of the cell sorter that generates harmful particles without enclosing the portion of the cell sorter that does not generate harmful particles, so as to minimize the size of the protected area and thus minimize the size of the integrated biological protection system 100 for the cell sorter. As Figure 1 shown, the nozzle chamber 134 contains a nozzle 146 that is supplied with a sample liquid 143 and a sheath liquid 145. The nozzle 146 generates a nozzle flow 153 ( Figure 2 ) from the sample liquid 143 and the sheath liquid 145, and the nozzle flow passes through the detection point 148 and through an opening 152 in the optical device mounting plate 150. Since the sample 143 may contain hazardous substances, such as hazardous cells that may be dispersed in aerosol form when the nozzle 146 is blocked, the nozzle 146 is contained within the nozzle chamber 134 to prevent any hazardous aerosol from escaping the aerosol management system chamber 140. The optical device mounting plate 150 separates the nozzle chamber 134 from the sorting chamber 131. The opening 152 allows the droplet flow 155 ( Figure 2 ) being detected to pass through the optical device mounting plate 150 to reach the sorting plate 154. Each droplet in the droplet flow 155 ( Figure 2 ) is detected at the detection point 148 and then separated by the sorting plate 154. The deflected droplet flow 156 is then collected by a collection medium 158. This is explained in more detail in U.S. Patent 8,557,587, issued to Fox et al. on October 15, 2013, the entire disclosure and teachings of which are specifically incorporated herein by reference.

[0013] The main cabinet protection system 101 is mainly used to control the sample input area 107 of the ambient air from outside the main cabinet protection system 101 ( Figure 5)。The sample input area 107 is located at the lower part of the main cabinet protection system 101. Samples are placed in the sample input area 107, which is part of the working area 104. The samples may include biohazardous substances. They are first prepared in a protected area, such as a large dedicated biological safety cabinet independent of the biohazard-protected cell sorter system disclosed herein. The cell samples are suspended in water and then capped to reduce the risk of contamination when the cell samples are removed from the large dedicated biological safety cabinet and transported to the integrated biohazard-protected cell sorter of the present invention. This reduces the risk that these cell samples may be contaminated by unwanted foreign substances during transportation to the integrated biohazard-protected cell sorter. Since the cells are suspended in water, the risk of accidental exposure to the user is very low. Even when the lid on the sample medium is removed, the risk of user exposure is low because the cells are not in aerosol form but are suspended in water. However, the user should wear safety glasses and gloves to reduce the risk of exposure due to accidental splashing of the sample into the eyes or mouth. Once the user places the capped sample tube in the sample input area 107, the tube cap can be opened in the clean environment of the sample input area, so the possibility of the sample being contaminated is very low. The tube is then placed in the sample input holder to start the sorting process.

[0014] Therefore, Figure 1 the shown main cabinet protection system 101 can intercept dangerous cells in the sample input area 107. As Figure 1 shown, when the partition 114, which may include a transparent sliding sash, is in the upper position in the access opening 115, the partition 114 allows the operator to easily enter the sample input area and the sorting chamber 131 through the sorting chamber door 132 to add and remove samples in the sample input area 107 ( Figure 5 ). The partition 114 can be moved to the lower position in the access opening 115 to allow the operator to directly enter the nozzle chamber 134 through the nozzle chamber door 136. The fan 122 is strong enough to maintain a low pressure in the working area 104 even when the partition 114 only covers part of the access opening. The partition 114 can simply move up and down in the access opening 115. Therefore, regardless of the position where the partition 114 is placed in the access opening 115, the amount of the area in the access opening 115 that is enclosed or blocked by the partition 114 and the amount of the area in the access opening 115 that is open and not enclosed or blocked by the partition 114 do not change. In other words, regardless of the position of the partition 114 in the access opening 115, the opening area of the access opening 115 is the same. As Figure 1As shown, when the baffle 114 is in the upper position, there is a certain number of square inches of opening in the access opening 115. When the baffle 114 is moved downward, the amount of the opening area in the access opening 115 has a constant size, that is, the number of square inches of the opening is the same, because the size of the baffle 114 is constant and the size of the access opening 115 is constant. Thus, no matter where the baffle 114 is located, the amount of air delivered by the fan 122 can be kept the same, and a constant first low pressure is still maintained in the working area 104. In one embodiment, the fan 122 moves about 100 feet of air per minute through the user access opening 115 into the working area 104. Functionally, the integrated biocontainment cell sorter system is based on the speed of air passing through the cabinet. The speed of the air must be fast enough to maintain containment because the integrated biocontainment cell sorter is not a sealed system. During operation, the speed of air entering the grate 118 is measured to ensure that the speed is appropriate to maintain containment. The fan 122 is designed to operate such that the amount of air passing through the grate 118 is sufficient to maintain containment of the harmful substances in the main cabinet protection system 101.

[0015] Figure 2 is a schematic diagram showing a part of the aerosol management system (AMS) 149. Figure 2 Specifically shown is a part of the cell sorter contained within the aerosol management system chamber 140. The nozzle 186 and connecting pipe fittings are located in the nozzle chamber 134. The sorting plate 176, collection tubes 182, 184 are located in the sorting chamber 131. The opening 174 allows the droplet stream 155 to flow through the optical device mounting plate 150 ( Figure 3 ) from the nozzle chamber 134 to the sorting chamber 131. Figure 2 Shows the main components located within the nozzle chamber 134 and the sorting chamber 131 including the aerosol management system chamber 140. Figure 2 Also schematically shows the main cabinet protection system 101. As Figure 2 shown, the nozzle 186 and connecting hose are located in the nozzle chamber 134. The sheath fluid container 170 contains sheath fluid, and the sheath fluid is delivered to the nozzle 186 through the sheath fluid hose 171. The sheath fluid hose 171 passes through the walls of the main cabinet protection system 101 and the aerosol management system chamber 140. The protective seal between the sheath fluid hose 171 and the walls of the main cabinet protection system 101 and the aerosol management system chamber 140 is not a protective seal that provides an airtight seal for the sheath fluid hose 171. Instead, a cheaper and easier-to-install seal can be used because both the main cabinet protection system 101 and the aerosol management system chamber 140 have a low pressure that causes air flow to enter inward into the main cabinet protection system 101 and the aerosol management system chamber 140. The same is true for the sample fluid hose 173.

[0016] Similarly as Figure 2As shown, the aerosol management system chamber 140 surrounds the sorting plate, collection tubes 182, 184, and the deflected droplet stream 156. The nozzle chamber 134 and the sorting chamber 131, as well as the opening 174 therebetween, include the aerosol management system chamber 140. The nozzle 186, the opening 174, the sorting plate 176, and the collection tubes 182, 184 are the main functional components contained within the aerosol management system chamber 140. The sample liquid container 172 placed in the sample input area 107 ( Figure 5 ) leads to the main cabinet protection system 101, rather than the aerosol management system chamber 140. The sheath liquid container 170, the excitation optics 162, the forward scatter detector 180, and the side scatter detector 178 are preferably all located outside the main cabinet protection system 101. As Figure 4 shown, the side scatter optical path 168 and the forward scatter optical path 166 project light through the optical windows onto the side scatter detector 178 and the forward scatter detector 180, respectively. Other electronics and controllers, as well as the laser, are preferably located outside the main cabinet protection system 101. For example, as disclosed in U.S. Patent 8,557,587, which is specifically incorporated herein by reference in its entirety for all of its disclosure and teachings, the timing and charging circuits, sorting logic controllers, filters, detectors, acquisition electronics, and other electronic circuits and devices, collectively referred to herein as the cell sorter electronics and optical equipment, are preferably all located outside the main cabinet and the aerosol management protection zone for ease of maintenance and adjustment. In other words, the cell sorter electronics and optical equipment are preferably located in an easily accessible area and do not require access to the contaminated areas within the main cabinet protection system 101 or the aerosol management system chamber 140. Thus, it is preferably possible to easily access the excitation optics 162, the side scatter detector 178, and the forward scatter detector 180 without accessing the biohazard area. Generally, these devices require adjustment, and the ability to access these devices without entering a dirty or biohazard area greatly improves the speed and maintenance of the system.

[0017] Most importantly in terms of simplifying maintenance and reducing the size of the protection system is to locate the excitation optics 162 outside the main cabinet protection system 101 and the aerosol management system 149. The excitation optics 162 includes an excitation laser or other excitation optics, such as an LED, a light-pumped plasma light generator, an arc lamp, or other excitation optics. The optics includes various mirrors, beam combiners, lenses, etc. Thus, according to one embodiment, the present invention can simply locate the excitation optics 162 outside the main cabinet and other components, such as the side scatter detector 178, the forward scatter detector 180, and other devices located inside the main cabinet 101 and the aerosol management system chamber 140. However, according to another embodiment of the present invention and as Figure 2As shown, optical detection devices such as the side scatter detector 178 and the forward scatter detector 180 can also be located outside the main cabinet 101 together with the excitation optics 162. As a third embodiment of the present invention, fluid devices such as the sheath fluid container 170 and various pumps associated with the sheath fluid can also be located outside the aerosol management system chamber 140 or the main cabinet protection system 101, either individually or together. In other words, various combinations of equipment can be located outside the main cabinet 101 and / or the aerosol management system chamber 140 to increase accessibility and ease of maintenance of the various systems and to reduce the size of the protected area.

[0018] Furthermore, the volume of the main cabinet protection system 101 is significantly reduced by creating an integrated system that is specifically constructed such that only the nozzle and the connecting hoses are in the nozzle chamber 134, and the sorting plate 176 and the collection tubes 182, 184 are in the sorting chamber. Many cell sorter systems are simply placed in very large and bulky enclosures. For example, the enclosures are typically about nine feet tall and can be six or seven feet wide. In the present invention, by enclosing only specific components, the protected area can be significantly reduced, and the overall size of the integrated biological protection system 100 can also be significantly reduced. By including the sample fluid container 172 in the input sample area 107 adjacent to the working area 104 of the main cabinet protection system 101, the input fluid can be easily added to and removed from the input sample area 107 because the sample fluid container 172 does not pose a danger like the aerosols generated in the nozzle chamber 134 and the sorting chamber 131. In addition, the use of the aerosol management system chamber 140 with an opening leading to the main cabinet protection system 101 provides additional safety for the operator because they are not affected by any harmful particles contained in the aerosols in the aerosol management system chamber 140.

[0019] Figure 3 is a schematic side cross-sectional view of the aerosol management system chamber 140. As Figure 3As shown, the nozzle chamber 134 contains a nozzle 186, which is shown in solid lines in the operating position. Sheath fluid 170 and sample fluid 172 are provided through the nozzle 186. The nozzle 186 can also be moved to a cleaning position, as shown by the dashed lines. The nozzle chamber door 198 can be opened so that the nozzle 186 in the cleaning position can be accessed through the nozzle chamber door 198. The nozzle, such as the nozzle 186, may become blocked for various reasons, and the nozzle 186 can be quickly and easily accessed when it is in the cleaning position. Similarly, when the nozzle chamber door 198 is open, the negative pressure in the nozzle chamber 134 is equalized compared to the main working area 104, so that contaminated air can migrate from the nozzle chamber 134 to the main working area 104 at that time point. The droplet stream 155 from the nozzle 186 flows through the opening 152 and the optical device mounting plate 150. The droplet stream 155 passes through the detection point 148 before the droplets are separated from the stream. The laser beam detects the nozzle stream 153 at the detection point 148. The scattered light and transmitted light from the detection point 148 pass through the optical window 200 and reach the side scatter objective 204 through the light blocking strip 202. The side scatter objective 204 collects the side scatter light rays and transmits these light rays through the side scatter aperture 206. The flexible seals 210, 212, and 214 provide a partial seal so that the droplet stream 155 does not transfer to the uncontrolled air 208. The droplet stream 155 passes through the sorting plate 154 and is separated into a deflected droplet stream 156. As Figure 1 shown, the AMS intake pipe 138 is connected to the AMS HEPA filter 141 and the AMS fan 144. Air from the sorting chamber 131 is drawn in through the AMS intake pipe 138 to create a low pressure in both the sorting chamber 131 and the nozzle chamber 134. Air from the main cabinet working area 104 passes through the inlets 188 and 194. The air stream 190 from the main cabinet passes through the inlet 188, while the air stream 196 from the main cabinet passes through the inlet 194. The sorting chamber 131 has a sorting chamber door 132, which can be opened to allow the operator to access the sorting plate 154 and the collection tubes 182, 184 in the sorting chamber 131 ( Figure 2 ). Similarly, since the pressure in the sorting chamber 131 is lower than that in the main cabinet working area 104, opening the sorting chamber door 132 equalizes the pressure between the sorting chamber and the main cabinet working area 104. To prevent harmful aerosols from escaping from the sorting chamber 131 or the nozzle chamber 134, the AMS fan 144 must be used ( Figure 1 ) to evacuate the sorting chamber 131 and the nozzle chamber 134 before opening the nozzle chamber door 198 or the sorting chamber door 132. Once the aerosols are cleared from the aerosol management chamber 140, the doors 198, 132 can be opened.

[0020] Figure 4 is a front view schematic diagram showing the nozzle chamber 134, the sorting chamber 131, and various devices of the cell sorter located outside the aerosol management system (AMS) 149 and the main cabinet protection system 101. As Figure 4As shown, uncontrolled air 208 surrounds the nozzle chamber wall 213 of the nozzle chamber 134. The laser 216 generates a laser beam along the optical path 164, and the laser beam passes through the optical window 215. The laser intersects the nozzle flow 153 at the detection point 148. The light transmitted from the monitoring point 148 passes through the optical window 217, reaches the light baffle 218, and passes through the forward scattering objective lens 220. The forward scattering objective lens 220 collects the light and transmits it through the pinhole 222 to the light detector 224. The droplet flow 155 passes through the opening 152 in the optical device mounting plate 150. The flexible seals 228 and 230 seal the sorting chamber 131 to the optical device mounting plate 150. The droplet flow 155 passes through the sorting plate 154 in the sorting chamber 131 and is separated by the sorting plate 154 into a deflected droplet flow 156. The sorting chamber wall 226 separates the sorting chamber 131 from the uncontrolled air region 208.

[0021] Figure 5 is a perspective view of an embodiment of a specific implementation of the integrated biocontainment cell sorter 100. As Figure 5 shown, the exhaust port 110 discharges clean air from the integrated biocontainment cell sorter 100. The clean air is supplied by the recirculation duct 108 to the recirculation plenum 102. The filter and fan plenum 121 houses the filter and fan in a position at the rear and bottom of the integrated biocontainment cell sorter 100, such that the operator can easily access other parts of the integrated biocontainment cell sorter 100. The sliding sash (partition) 114 moves in an angled vertical motion to provide an opening to the main cabinet protection system 101. As shown, when the sliding sash 114 is in the upper position, the operator can access the sample input area 107. When the sliding sash 114 is in the lower position, access to the nozzle chamber 134 and the sample line is available. The grate 118 allows air from outside the integrated biocontainment cell sorter 100 to be drawn into the main cabinet protection system 101 so that contaminated air does not flow out of the main cabinet protection system 101. By maintaining a lower pressure in the main cabinet protection system 101, contaminated air does not escape from the integrated biocontainment cell sorter 100.

[0022] Therefore, the integrated biocontainment cell sorter 100 provides protection only around the parts of the cell sorter where contaminated air may be generated. Therefore, compared with a cell sorter placed in a large hood or a partially integrated biocontainment cell sorter, the integrated biocontainment cell sorter 100 has a smaller and more compact protection area, a smaller fan, and a significantly smaller overall size. The partially integrated biocontainment cell sorter encapsulates many components in the cell sorter that do not require protection, and it is difficult to maintain specific areas of the cell sorter that do not require biocontainment. The smaller protection area results in a smaller fan and a reduced amount of air that needs to be moved to maintain the protection, thereby saving energy.

[0023] The foregoing description of the present invention has been presented for purposes of illustration and description. The foregoing description is not intended to be exhaustive or to limit the invention to the precise form disclosed, and many modifications and variations are possible in light of the above teachings. The embodiments were chosen and described in order to best explain the principles of the invention and its practical application, so as to enable others skilled in the art to best utilize the invention in various embodiments and various modifications as are suited to the particular use contemplated. The appended claims are intended to be construed to include alternative embodiments of the present invention, except as limited by the prior art.

Claims

1. An integrated bioprotective cell sorter flow cytometer, comprising: A main cabinet of the integrated bioprotective cell sorter, which is not airtight; An input sample area, which is arranged inside the main cabinet; A movable partition, which is located inside the main cabinet and moves in the entrance and exit of the main cabinet; when the movable partition moves in the entrance and exit, the movable partition covers a constant area of the entrance and exit, which makes the area not covered by the movable partition in the entrance and exit remain constant and the area not covered by the movable partition is open when the movable partition moves in the entrance and exit; A first fan, which sucks air from the main cabinet to generate a first low pressure in the main cabinet, and the first low pressure is basically constant when the movable partition moves in the entrance and exit, which restricts the contaminated air in the main cabinet from escaping from the main cabinet; An aerosol management protection area, which is not airtight and is arranged in the main cabinet; the aerosol management protection area has an opening, and the opening is connected to the main cabinet so that the aerosol management protection area is arranged in the first low pressure and is subjected to the first low pressure; The aerosol management protection area has a nozzle for generating a droplet stream containing sample cells, a sorting plate for dividing the droplet stream into deflected streams, and a collection medium for collecting the deflected streams; A second fan, which sucks air from the aerosol management protection area to generate a second low pressure lower than the first low pressure in the aerosol management protection area, so that the air from the main cabinet flows from the main cabinet to the aerosol management protection area and restricts the contaminated air from flowing into the main cabinet from the aerosol management protection area; A light excitation device, which is located outside the main cabinet and the aerosol management protection area to allow access to the light excitation device without entering the main cabinet or the aerosol management protection area.

2. The integrated bioprotective cell sorter flow cytometer according to claim 1, wherein The movable partition is a sliding window.

3. The integrated bioprotective cell sorter flow cytometer according to claim 1, wherein the light excitation device comprises a laser and excitation optics.

4. The integrated bioprotective cell sorter flow cytometer according to claim 3, wherein the detector optics, filters, timing and charging circuits, and sorting logic controller are located outside the main cabinet.

5. The integrated bioprotective cell sorter flow cytometer according to claim 3, wherein the fluid device is located outside the main cabinet.

6. The integrated bioprotective cell sorter flow cytometer according to claim 5, wherein the fluid device comprises a sheath fluid controller and a nozzle cleaning device.

7. The integrated bioprotective cell sorter flow cytometer according to claim 2, further comprising: A first HEPA filter, which is located near the main working area of the main cabinet and upstream of the first fan, so that the first fan circulates clean air and is not contaminated; A second HEPA filter, which is located near the aerosol management protection area and upstream of the second fan, such that the second fan is not contaminated and removes the air cleaned by the second HEPA filter from the aerosol management protection area, and moves the clean air to the exhaust port.

8. A method for intercepting cells in an integrated biological protection cell sorter, comprising: Providing a main cabinet protection area, which contains an input area for the sample cells to be sorted; Using a first fan to generate a first low pressure in the main cabinet, the first fan sucking air from the main cabinet and sucking air from outside the cabinet; Using a second fan to generate a second low pressure in an aerosol management protection area provided in the main cabinet, the second fan sucking air from the aerosol management protection area through an opening connected to the main cabinet in the aerosol management protection area to generate the second low pressure lower than the first low pressure in the aerosol management protection area; Enclosing the input cell sample in the main cabinet that is not airtight sealed; Enclosing the nozzle, sorting plate, collection medium, and any droplet stream generated by the nozzle in the aerosol management protection area that is not airtight sealed; Placing the photoexcitation device outside the main cabinet and the aerosol management protection area to facilitate adjustment and maintenance of the photoexcitation device.

9. The method according to claim 8, wherein the method of placing the photoexcitation device outside the main cabinet includes placing the laser and laser optics outside the main cabinet.

10. The method according to claim 8, further comprising: Placing the detector optics, filter, timing and charging circuits, acquisition electronics, and sorting logic controller outside the main cabinet.

11. The method according to claim 8, further comprising: Placing the fluid device outside the main cabinet.

12. The method according to claim 11, wherein the method of placing the fluid device outside the main cabinet includes: Placing the sheath fluid controller and nozzle cleaning device outside the main cabinet.

13. The method according to claim 8, further comprising: Using a movable partition in the entrance and exit of the main cabinet, the movable partition covering a certain area of the entrance and exit such that when the movable partition moves in the entrance and exit, a constant area of the entrance and exit is not covered by the movable partition and the area not covered by the movable partition remains constant, which results in the first low pressure remaining constant when the movable partition moves in the entrance and exit.

14. The method according to claim 13, wherein the movable partition is transparent and moves to allow entry into the lower part of the main cabinet when the movable partition is in the upper position, and moves to allow entry into the upper part of the main cabinet when the movable partition is in the lower position.

15. The method according to claim 8, further comprising: Air inhaled outside the filtration cabinet upstream of the first fan and air inhaled from the main cabinet are filtered so that the first fan is not contaminated, and the air under the pressure from the first fan is clean air that can circulate in the integrated biological protection cell sorter without being contaminated.

16. The method according to claim 8, further comprising: Filtering the air drawn from the aerosol management protection area upstream of the second fan so that the second fan is not contaminated, and the clean air under the pressure from the second fan is conveyed through the integrated biological protection cell sorter and discharged without contamination.

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