Introduction
-Bacstain- series offer several kinds of bacterial fluorescence staining dye and can be applied for microbial cell viability assay in different principles.
DAPI is an AT-sequence specific DNA intercalator that attaches to DNA at the minor groove of the double helix.
-Bacstain-DAPI solution is a Ready-to-Use solution.
Kit contents
DAPI aqueous solution | 25 μl x 4, 1 mg/ml |
Storage
Store at 0-5°C.
Required Equipment
- Fluorescence microscope (ultraviolet excitation light, blue emission filter) or Flow cytometer (ultraviolet light, blue emission filter)
- Micropipette (20 μl, 1,000 μl)
Precaution
This kit includes microtubes containing solutions. Since there is a possibility that the droplets might attach to the inside walls or caps, please shake them down prior to open.
Sraining Procedure
- Allow DAPI solution to stand at room temperature for 30 minutes. Solution should be protected from light a).
- Resuspend the organisms with PBS(-) or saline and adjust the number of cells to 106 cells/ml (flow cytometry) or 108 -109 cells/ml (microscopy).
- Add 1μl of DAPI solution into the 1 ml of microbial cell suspension and vortex gently to mix.
- Incubate the microbial cells at room temperature for 5 minutes.
- Analyze the stained cells by flow cytometer or under a microscope.
a) Since DAPI may be carcinogenic, please be careful in its handling/disposing.
- DAPI-stained B.cereus
- Left: Fluorescence image , Right: Bright field image
Number of Tests Possible
This kit can perform at least 100 tests for the flow cytometric assay and microscopic assay by following the protocol described herein.
Doublestaining (Optimal)
If -Bacstain- CTC Rapid Staining Kit or -Bacstain- CFDA solution is used with DAPI, double-staining examination can be performed.
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L.casei were stained with CTC, and then counterstained with DAPI.
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B.cereus were stained with CFDA, and then counterstained with DAPI.
Relevant Products
-Bacstain- CTC Rapid Staining Kit (for Flow cytometry)
-Bacstain- CTC Rapid Staining Kit (for Microscopy)
CTC has been used by many researchers to evaluate the microbial respiratory activity.
-Bacstain- CTC Rapid Staining Kit allows quick and high-sensitivity CTC-staining.
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Right-image : without Enhancing reagent
Left-image : with Enhancing reagent
Ex filter : Blue
Em filter: Red
CTC staining efficiencies were compared in with or without enhancing reagent condition.
-Bacstain- CFDA solution
CFDA is widely used as an indicator for the measurement of esterase activity.
-Bacstain- CFDA solution is provided as Ready-to-Use DMSO solution.
Fluorescent carboxyfluorescein is produced from non fluorescent CFDA by the esterase in the microbial cell.
-Bacstain- AO solution
AO is nucleic acid staining dyes and can be applied for bacteria either with membrane impairment or with membrane intact.
-Bacstain- PI solution
PI is a nucleic acid staining dye. Membrane-injured cells are stained with red emission by PI.
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Double-staining of S.epidermidis (DAPI/PI)
Red fluorescence represents membrane-injured cells
Products | Code | Maximum Ex/Em(nm) | Number of assays |
CTC Rapid Staining Kit (for Flow cytometry) | BS01 | 430, 480/630 | 100 |
CTC Rapid Staining Kit (for Microscopy) | BS02 | 430, 480/630 | 100 |
CFDA solution | BS03 | 493/515 | 100 |
DAPI solution | BS04 | 360/460 | 100 |
AO solution | BS05 | 420-460/630-650(ssDNA) | 100 |
500/520(dsDNA) | |||
PI solution | BS07 | 530/620 | 100 |
Frequently Asked Questions / Reference
BS04: -Bacstain- DAPI solution
Revised Aug., 23, 2023