Multi-fluorescent staining solution as well as preparation method and use method thereof

A multi-fluorescence and dyeing solution technology, applied in the field of multi-fluorescence detection, can solve the problems of low dyeing clarity of the target object, high requirements for imaging equipment configuration, unclear imaging outline, etc., achieving clear effect, short operation time, and clear boundary. Effect

Pending Publication Date: 2021-10-22
SHENZHEN UNI MEDICA TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Each sample needs to be coated with two glass slides, one slide is stained with fungi, and is detected by UV fluorescence module; the other slide is stained with microorganisms and other formed components other than fungi, and is detected by blue light fluorescence module; The detection method has long operation time, cumbersome steps, and requires two fluorescence modules, which requires high configuration of imaging equipment
[0003] And at present, the dyeing clarity of similar products to the target is not high, especially the imaging outline of small targets (such as bacteria, fungi, etc.) is not clear, which will cause errors (omissions and mistakes) in the recognition (manual and automatic) process recognition); the current traditional non-staining method (salt water sheet detection) recognition accuracy, due to the limitations of the technology itself, can only reach a maximum of 75%-80%

Method used

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  • Multi-fluorescent staining solution as well as preparation method and use method thereof
  • Multi-fluorescent staining solution as well as preparation method and use method thereof
  • Multi-fluorescent staining solution as well as preparation method and use method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0043] Prepare multiple fluorescent staining solutions, including the following steps:

[0044] S1: Weigh or measure the following substances: NaCl 100mM, KCl 1mM, NaCl 2 HPO 4 0.5mM, KH 2 PO 4 5mM, mouse anti-vaginal Gardner monoclonal antibody A / B / C 1g, rabbit anti-Candida albicans monoclonal antibody A / B 1g, mouse anti-N. gonorrhoeae monoclonal antibody A / B 1g, mouse anti-vaginal Trichomonas monoclonal antibody A / B 1g, mouse anti-herpes simplex virus (type I / II) monoclonal antibody A / B 1g, Proclin300 0.01L, DMSO 0.1L, Tween 20 0.001L, Hochest 0.01mg, 4, 4'-Bis(2-benzoxazolyl)stilbene 0.001mg, acridine orange 0.001mg, carboxyfluorescein diacetate succinimidyl ester 0.001mg, Sybr Green I 0.001mg, Sybr GreenⅡ0.001mg , cell membrane fluorescent probe (DiI) 0.0001mg, cell membrane fluorescent probe (DiO) 0.0001mg, cell membrane fluorescent probe (DiR) 0.0001mg, fluorescein isothiocyanate 0.001mg, phycoerythrin 0.001mg, peridinoxanthin Phyllite-chlorophyll-protein complex ...

Embodiment 2

[0050] Prepare multiple fluorescent staining solutions, including the following steps:

[0051] S1: Weigh or measure the following substances: NaCl 135mM, KCl 2.7mM, NaCl 2 HPO 4 1.5mM, KH 2 PO 4 8mM, mouse anti-vaginal Gardner monoclonal antibody A / B / C 5g, rabbit anti-Candida albicans monoclonal antibody A / B 10g, mouse anti-N. gonorrhoeae monoclonal antibody A / B 7.5g, mouse anti- Trichomonas vaginalis monoclonal antibody A / B 12g, mouse anti-herpes simplex virus (I / II type) monoclonal antibody A / B 6.7g, Proclin300 0.05L, DMSO 2.5L, Tween 20 0.01L, Hochest0.05mg, 4,4'-bis(2-benzoxazolyl)stilbene 10mg, acridine orange 10mg, carboxyfluorescein diacetate succinimide ester 50mg, Sybr Green I 10mg, Sybr GreenⅡ10mg, cell membrane fluorescence probe Needle (DiI) 10mg, cell membrane fluorescent probe (DiO) 10mg, cell membrane fluorescent probe (DiR) 10mg, fluorescein isothiocyanate 10mg, phycoerythrin 10mg, peridinoxanthin-chlorophyll-protein complex 10mg, Propidium iodide 10mg,...

Embodiment 3

[0057] Prepare multiple fluorescent staining solutions, including the following steps:

[0058] S1: Weigh or measure the following substances: NaCl 200mM, KCl 5mM, NaCl 2 HPO 4 2mM, KH 2 PO 4 158mM, mouse anti-vaginal Gardner monoclonal antibody A / B / C 50g, rabbit anti-Candida albicans monoclonal antibody A / B 50g, mouse anti-N. gonorrhoeae monoclonal antibody A / B 50g, mouse anti-vaginal Trichomonas monoclonal antibody A / B 50g, mouse anti-herpes simplex virus (type I / II) monoclonal antibody A / B 50g, Proclin300 0.2L, DMSO 50L, Tween 20 20L, Hochest 10mg, 4,4'- Bis(2-benzoxazolyl)stilbene 0.02mg, acridine orange 0.02mg, carboxyfluorescein diacetate succinimide ester 0.05mg, Sybr Green I 0.01mg, Sybr GreenⅡ0.01mg, cell membrane fluorescence Probe (DiI) 0.001mg, cell membrane fluorescent probe (DiO) 0.001mg, cell membrane fluorescent probe (DiR) 0.001mg, fluorescein isothiocyanate 0.01mg, phycoerythrin 0.01mg, peridinoxanthin-chlorophyll -Protein complex 0.01mg, propidium iod...

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Abstract

The invention discloses a multiple fluorescent staining solution as well as a preparation method and a use method thereof. Each liter of the multiple fluorescent staining solution is prepared from the following components: 105.5-365 mM of strong base, 0.01-50 g of acid, 5-250 g of binding protein, 0.01-0.2 L of preservative, 0.1-50 L of solubilizer, 0.001-20 L of membrane rupture agent, 0.0194-180 mg of fluorescent staining agent and the balance of sterilized distilled water. According to the method, fungus detection substances and detection substances of other microorganisms except fungi are chelated together, the detection result can be obtained only through one-time smearing, one-time dyeing and one-time color observing, the effect is clearer after dyeing, the dyeing boundary of a target object is more sharpened, and therefore the method is very beneficial to small target object recognition; particularly, bacteria and fungi in microorganisms have small forms, clear boundaries and a sharpened fluorescent staining effect, so that the identification of the target object is facilitated.

Description

technical field [0001] The invention belongs to the technical field of multiple fluorescence detection, in particular to the detection of gynecological vaginal microecology; in particular, it relates to a multiple fluorescence staining liquid and its preparation method and use method. Background technique [0002] The existing gynecological vaginal microecology detection method uses a two-step method to stain the samples, that is, the detection of fungi in each sample must be stained separately. Each sample needs to be coated with two glass slides, one slide is stained with fungi, and is detected by UV fluorescence module; the other slide is stained with microorganisms and other formed components other than fungi, and is detected by blue light fluorescence module; The detection method has long operation time, cumbersome steps, and requires two fluorescence modules, which requires high configuration of imaging equipment. [0003] And at present, the dyeing clarity of similar...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N1/30G01N21/64
CPCG01N1/30G01N21/6428G01N21/6458G01N2001/302G01N2021/6439
Inventor 郭永超蔡锦刚王艳平黄萍
Owner SHENZHEN UNI MEDICA TECH
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