Claims
- 1. A method for quantifying the relative number of viable cells in a cell sample, comprising:treating the sample with a first, fluorescent dye, wherein the first dye substantially accumulates in viable cells only; treating the sample with a second dye, wherein the second dye quenches the fluorescence of the first dye, and wherein the second dyepenters non-viable cells but is substantially excluded by viable cells; and measuring an intensity of fluorescence of the sample.
- 2. The method of claim 1, wherein the first dye is an ester of fluorescein.
- 3. The method of claim 1, wherein the second dye is an acid dye.
- 4. The method of claim 1, wherein the first dye is fluorescein diacetate or related dyes and the second dye is eosin Y.
- 5. The method of claim 1, wherein the first dye is calcein-AM and the second dye is trypan blue.
- 6. The method of claim 1, wherein the second dye reduces the fluorescence of the first dye by being in close proximity to the first dye.
- 7. The method of claim 1, wherein the measuring step comprises:obtaining at least one digital image of a defined area of the container, the digital image comprising pixels and representing fluorescence light intensities at each pixel; and extracting from the digital image information relating to relative cell number.
- 8. A method of preparing a cell sample for quantification of cell numbers using fluorescence digital imaging microscopy, comprising:treating the sample with a first, fluorescent dye, wherein the first dye substantially accumulates in viable cells only; and treating the sample with a second dye, wherein the second dye quenches the fluorescence of the first dye, and wherein the second dye enters non-viable cells but is substantially excluded by viable cells.
- 9. The method of claim 8, wherein the first dye is an ester of fluorescein.
- 10. The method of claim 8, wherein the second dye is an acid dye.
- 11. The method of claim 8, wherein the first dye is fluorescein diacetate and the second dye is eosin Y.
- 12. The method of claim 8, wherein the first dye is calcein-AM and the second dye is trypan blue.
- 13. The method of claim 8, wherein the second dye reduces the fluorescence of the first dye by binding to the first dye.
Parent Case Info
This is a continuation-in-part of application Ser. No. 08/622,110, filed Mar. 26, 1996.
US Referenced Citations (10)
Non-Patent Literature Citations (1)
Entry |
Tanz et al., “Further Studies on Selection for Viable Cells in Culture Using Cell Sort Technology”, Proceedings of Annual International Conference of the IEEE, 1988, pp. 303-304. |
Continuation in Parts (1)
|
Number |
Date |
Country |
Parent |
08/622110 |
Mar 1996 |
US |
Child |
09/066134 |
|
US |