Cytological and histological fixative composition and methods of use
A composition and histological technique, applied in biochemical equipment and methods, immobilized enzymes, scientific instruments, etc.
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Embodiment 1
[0066] Embodiment 1. fixative preparation
[0067] The following fixative compositions were produced: 40% alcohol (methanol and isopropanol); 2.2% acetone; 1.8% polyethylene glycol (PEG); 0.7% formaldehyde; 1% glycerin; Buffer (pH7.4-7.8). All ingredient amounts are about percent by volume.
[0068] The fixative solutions described above are used in conjunction with various cytology specimen preparation techniques or devices, including those described in U.S. Patent 5,471,994; U.S. Patent 5,301,685; U.S. Serial No. (USSN) 08 / 905,833 filed Aug. USSN 08 / 963,873 filed on March 13, 1998, USSN 09 / 053,010 filed on April 1, 1998; PCT / US98 / 16349; USSN 09 / 050,010; PCT / US98 / 17524 ; those disclosed in USSN 09 / 185,606 and PCT / US98 / 23222. The fixative solution of Example 1 demonstrated rapid tissue penetration with a minimal amount of preparation; tissue stained in this way showed excellent histological and cytological detail.
Embodiment 2
[0069] Example 2. Cytological preparation
[0070] 1. The specimen collection container is half-filled with the fixative composition of the present invention. Place the specimen containing the cells into the container with the fixative composition.
[0071] 2. Mix the cytology specimen and fixative composition vigorously so that the specimen and fixative are thoroughly mixed.
[0072] 3. If the cells of the cytology specimen are not dispersed, the specimen is probably homogeneous. Homogenization is performed with a mechanical blade rotating at about 2,000 to 4,000 revolutions per minute (RPM), preferably at about 3,000 RPM. MonoGen TM The blades at the bottom of the Monoprep G perform this function. Alternatively, beat with a high-speed mixer for 5 to 10 seconds, or less. If spots and fine lines are visible in the specimen, return it to the blender and blend for another 10 to 15 seconds. Avoid excessive stirring.
[0073] 4. Cells should be fixed for at least 15 min...
Embodiment 3
[0074] Example 3 Cytology Preservation and Mucolytic Solution
[0075] The following fixative and mucolytic compositions were produced: 40% alcohol (methanol and isopropanol); 2.2% acetone; 1.8% polyethylene glycol (PEG); 0.7% formaldehyde; 1% glycerol; 0.02% Nonidet P40; and 54% Tris buffer (pH7.4-7.8). All ingredient amounts are about percent by volume. DTT was added in the range of 0.2-0.5 gram%. The solution is called MonoLex TM . The MonoLex TM The solution was added to the specimen at a ratio of 1:1. Spin the specimen and mucolytic fixative solution for approximately 1 minute, then incubate for about 30 minutes, and spin again for about 1 minute. Further processing includes methods well known in the art for preparation of particle monolayers and staining of cells. Various cytological preparation devices, such as those listed in Example 1, can be used.
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