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Device for analysis of cellular motility

Inactive Publication Date: 2016-03-10
MOTILITYCOUNT
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Benefits of technology

The present invention is about a method for estimating the amount of motile cells in a sample. This is important because motile cells are important for various applications such as medical diagnosis and research. The method involves using a cell permeable filter to separate motile cells from non-motile cells. The application compartment is below the filter and the conditioning medium compartment is above the filter. This setup significantly improves the effect of the filter. The method is simple and can be carried out in a home environment without advanced technical knowledge.

Problems solved by technology

Some drawbacks with the prior art devices exist, which provide for a limited accuracy when estimating the number of motile cells.
In particular, the presently available devices are not capable of sufficiently reducing contamination such as by immotile cells, even when cell permeable filters are integrated.
In addition, the migration of motile sperm cells through fluidic channels may decrease the concentration at the point of quantification to a level which is difficult to detect.

Method used

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  • Device for analysis of cellular motility
  • Device for analysis of cellular motility
  • Device for analysis of cellular motility

Examples

Experimental program
Comparison scheme
Effect test

example 1

[0112]Experiments were conducted to compare the effect of having the sample application compartment below the cell permeable filter and the conditioning medium compartment above the cell permeable filter with previous applications of cell permeable filters, where the sample application compartment is e.g. beside or above the conditioning medium compartment.

[0113]A comparison between having sample application compartment above and below the cell permeable filter was carried out.

[0114]A semen sample containing only immotile cells was used. The sample was then split into two samples for testing.

[0115]Sample Application Compartment Above a Cell Permeable Filter

[0116]One sample was applied to a system having a sample application compartment above a cell permeable filter and a conditioning medium below the filter. The conditioning medium was pipetted into the conditioning medium compartment and the semen sample was pipetted into the sample application compartment. A sample of conditioning...

example 2

[0121]The dynamic range of the colour intensity of the RGB values of image data for an image from a mesoscale fluidic device of the invention was correlated with the manual count of a titrated semen sample. A semen sample containing 185 million motile sperms per millilitre was diluted 2-fold serially with semen plasma. The samples were applied to the test devices using standard incubation parameters using MTT at a concentration of 0.5 mg / ml as a detection agent. Photographs of the results were taken with a Microsoft life digital camera and the colour intensities (RGB values) measured. The mesoscale fluidic device comprised a cell retaining filter, and for each image for analysis the mesoscale fluidic device was placed in a light box to ensure the same conditions at all times. The light box consisted of a box and included the camera positioned approximately 55 mm from the object and a 12 volt halogen bulb. The camera attributes were controlled by a custom made Labview program. Condit...

example 3

[0123]The possible effects of two detection agents, MTT and crystal violet, on the motility of sperm cells were tested. Stock solutions of MTT and crystal violet of 5 mg / ml and 1 mg / ml, respectively, were prepared, and these were diluted two-fold serially. The detection agent solutions were mixed with sperm samples comprising 60 million motile cells per millilitre and 72 million cells per millilitre in total (denoted 60 / 72) in a 50:50 ratio and the mixtures were incubated for 60 min at ambient temperature before analysing the cells microscopically.

TABLE 3MTT (mg / ml)CellsCrystal violet (mg / ml)Cells060 / 72060 / 72518 / 64140 / 632.530 / 630.527 / 431.2534 / 650.2532 / 560.62548 / 770.12537 / 56

[0124]It is evident from Table 3 that MTT had an effect on the cellular motility. The ratio of motile versus non-motile cells (% motile) was lowered at the higher concentrations, whereas the ratio was not affected by crystal violet. However, at MTT concentrations below 2.5 mg / ml no effect on motility was observed ...

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Abstract

A mesoscale fluidic system comprises a substrate having a sample chamber and an analysis chamber. The sample chamber comprises a cell permeable filter defining a sample application compartment and a conditioning medium compartment. The sample chamber has a sample inlet port in the sample application compartment. The analysis chamber has an entry port and an exit port. The conditioning medium compartment is in fluid communication with the entry port of the analysis chamber via a channel. The sample application compartment is below the cell permeable filter and the conditioning medium compartment is above the cell permeable filter. The mesoscale fluidic system is suited for analysing cellular motility in a sample. Also disclosed is a method of estimating the quantity of motile cells in a sample and a method of extracting motile cells from non-motile cells.

Description

[0001]This invention relates to a mesoscale fluidic system for analysing cellular motility in a sample, in particular the motility of cells in a sperm sample. The present invention also relates to a method of estimating the quantity and quality of motile cells in a sample and to a method of extracting motile cells from non-motile cells, in particular using the mesoscale fluidic system.PRIOR ART [0002]Awareness of decrease in male fertility potential has been high during the last decades, thus creating a demand for having sperm samples analysed. In particular, it is known that the concentration of motile sperm cells in a sample is the most predictive factor with regard to estimating the fertility of a sample (Tomlinson et al., 2013, Human Fertility, 1-19). At present an analysis for assessing the number and motility of cells in a sperm sample is typically carried out by professionals in a laboratory environment, and home diagnostic devices for estimating male fertility potential have...

Claims

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Application Information

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IPC IPC(8): B01L3/00G01N1/40
CPCB01L3/502753G01N1/4077G01N2001/4088B01L2300/04B01L2400/0478B01L2300/0681G01N2015/0003C12M41/46B01L2300/087B01L2400/0487G01N15/01
Inventor LARSEN, JACOB MOLLENBACHLAURSEN, STEEN BROCH
Owner MOTILITYCOUNT
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