Dojindo,生物膜形成检测试剂盒/100/B601

2022年5月1日 作者 jinpanbio
生物膜是由微生物和细胞外多糖组成的生物聚集体,可以存在于许多环境中。 生物膜中的微生物对抗生素具有很强的抗药性; 因此,对具有抗生物膜活性的药物和食品成分的研究是一个不断发展的领域。我们的检测试剂盒用于定量测量生物膜形成/抑制生物膜形成(Biofilm Formation Assay Kit)和抗生物膜药物功效(Biofilm Viability Assay Kit) 采用了一种协议,该协议旨在使用来自生物膜形成的钉板。 我们的检测试剂盒将所有必要的成分组合到每个包装中。

Feature 1: Measurement time can be significantly shortened.In existing methods, biofilms are formed on the bottom of the well of a microplate, and additional work is required to change microbial culture media and wash sample several times before and after staining. However, our assay kits allow you to form biofilms on the pegs attached to the plate’s lid. Media change and staining process can be done simply by transferring the lid; therefore, our assay kits are very easy to use.

Feature 2: Our assay kits can reduce measurement variation.In existing methods, biofilm formation occurs on the bottom of the well of a microplate, which made biofilm easier to peel off because of the processes such as washing. This causes measurement variation, and this measurement variation was an issue for the quantification of biofilm. Our assay kits allow you to form biofilms on the pegs attached to the plate’s lid and make it possible to avoid peeling off biofilm which occurs during the process of the formation.

Two different types of assay kits for different purposes.We offer two types of assay kits that measure the amount of biofilm formation or the metabolic activities of live microorganisms within biofilms by using the same measurement method. You can select the one that best suits your needs.

Selection of 2 types of assay kits

*Conditions on biofilm formation vary according to microbial species and strains. When you need to examine these conditions, we first recommend you use the Biofilm Formation Assay Kit.*The Biofilm Formation Assay Kit is a product developed together with Fukuoka Industrial Technology Center.

Procedure

Example of measurementIndicators for drug-mediated inhibition of biofilm formation or antimicrobial effect of biofilms include MBIC (minimum biofilm inhibitory concentrations) and MBEC (minimum biofilm eradication concentrations). MBIC and MBEC of S. aureus were measured using each kit.

[How to use – Quick Tutorial]
Q1. How can I examine the conditions on biofilm formation?

A1. Test items for conditions on biofilm formation include types of culture media, seeding density of microorganisms, frequency of media changes, incubation time, incubation temperature, and others.We show examples of optimizing the following test items: types of culture media, seeding density of microorganisms, frequency of media changes, and incubation time, as below.If you optimize incubation temperature, it is necessary to use a number of the Biofilm Formation Assay Kits (Product code: B601).Measure the amount of biofilm formation – ①Under the following optimized conditions: types of culture media, seeding density of microorganisms, and media changes, the amount of biofilm formation can be confirmed by the experiments shown below.

Example of plate arrangement

Experimental schedule

Procedure(Step 1) Day 1: 96-well plate ①In accordance with the reference layout shown above, fill each well (rows A and B) of the 96-well plate with 180μl of microbial cell suspension with concentrations ① and ② made in culture media A through D.Leave the wells (rows C through H) blank. Place a 96-peg lid on the 96-well plate and incubate the plate at the optimum growth temperature for the microorganism for 24 hours.※ ExamplesMicrobial concentrations ① and ②: Approximately 107 CFU/ml and 106 CFU/ml, respectively.(Step 2) Day 2: 96-well ②In accordance with the reference layout shown above, fill each well (rows A and B) of the 96-well plate with 180μl of culture media without microorganism.Fill each well (rows C and D) of the same 96-well plate with 180μl of microbial cell suspension with concentrations ① and ② made in culture media A through D. Leave the wells (rows E through H) blank.Place the 96-peg lid (Step1) on the 96-well plate ② and incubate the 96-well plate ② at the optimum growth temperature for the microorganism for 24 hours.(Step 3) Day 3: 96-well plate ③In accordance with the reference layout shown above, fill each well (rows A through D) of the 96-well plate with 180μl of culture media without microorganism.Fill each well (rows E and F) of the same 96-well plate with 180 μl of microbial cell suspension with concentrations ① and ② made in culture media A through D. Leave the wells (rows G and H) blank.Place the 96-peg lid (Step1) on the 96-well plate ③ and incubate the 96-well plate at the optimum growth temperature for the microorganism for 24 hours.(Step 4) Day 4: 96-well plate ④In accordance with the reference layout shown above, fill each well (rows A through F) of the 96-well plate with 180μl of culture media without microorganism.Fill each well (rows G and H) of the same 96-well plate with 180μl of microbial cell suspension with concentrations ① and ② made in culture media A through D.Place the 96-peg lid (Step1) on the 96-well plate ④ and incubate the 96-well plate ④ at the optimum growth temperature for the microorganism for 24 hours.(Step5) Perform experiments using a new 96-well plate, in accordance with Steps 2 through 6 which describe the measurements of biofilm formation/inhibition of biofilm formation in the technical manual.Measure the amount of biofilm formation – ②Under the following optimized conditions: types of culture media, seeding density of microorganisms, and media changes, the amount of biofilm formation can be confirmed by the experiments shown below.

Example of plate arrangement

Experimental schedule

Procedure(Step 1) Day 1: 96-well plate○1In accordance with the reference layout shown above, fill each well (rows A and B) of the 96-well plate with 180μl of microbial cell suspension with concentrations ① and ② made in culture media A through D.Leave the wells (rows C through H) blank.Place a 96-peg lid on the 96-well plate and incubate the plate at the optimum growth temperature for the microorganism for 24 hours.※ ExamplesMicrobial concentrations ① and ②: Approximately 107 CFU/ml and 106 CFU/ml, respectively.(Step2) Day 2: Remove the 96-peg lid. Allow cells to grow in the rows A and B of the 96-well plate○1 without changing the culture medium.Fill each well (rows C and D) of the same 96-well plate with 180μl of microbial cell suspension with concentrations ① and ② made in culture media A through D. Leave the wells (rows E through H) blank.Place the 96-peg lid back to the 96-well plate ①. Continuously incubate the 96-well plate ① at the optimum growth temperature for the microorganism for 24 hours.(Step3) Day 3: Remove the 96-peg lid. Allow cells to grow in the rows (A through D) of the 96-well plate ①without changing the culture medium.Fill each well (rows E and F) of the same 96-well plate with 180μl of microbial cell suspension with concentrations ① and ② made in culture media A through D. Leave the wells (rows G and H) blank. Place the 96-peg lid back to the 96-well plate ①. Continuously incubate the 96-well plate ① at the optimum growth temperature for the microorganism for 24 hours.(Step 4) Day 4: Remove the 96-peg lid. Allow cells to grow in the rows (A through F) of the 96-well plate ① without changing the culture medium.Fill each well (rows G and H) of the same 96-well plate with 180 μl of microbial cell suspension with concentrations ① and ② made in culture media A through D. Leave the wells (rows G and H) blank. Place the 96-peg lid back to the 96-well plate ①. Continuously incubate the 96-well plate ① at the optimum growth temperature for the microorganism for 24 hours.(Step 5): Perform experiments using a new 96-well plate, in accordance with Steps 2 through 6 which describe the measurements of biofilm formation/inhibition of biofilm formation in the technical manual.

Q2. What types of microbial species have been tested?

A2. Dojindo Laboratories has evaluated the following microbial species that formed biofilms using the Biofilm Formation Assay Kit (this product) and the Biofilm Viability Assay Kit (Product code: B603).– Staphylococcus aureus– Pseudomonas aeruginosa– Escherichia coli– Streptococcus mutans– Porphyromonas gingivalisPlease refer to the technical manual, in terms of culture conditions for the above microbial species, experimental examples, etc.

Q3. When I evaluate inhibition of biofilm formation and anti-biofilm drug efficacy, what is the approximate amount of biofilm formation required for experiment?

A3. In the Biofilm Formation Assay Kit (Product code: B601), please consider the amount of biofilm formation as a rough indication when a value of absorbance (590 nm) for crystal violet solution obtained after the final step is greater than 0.5.

Dojindo细胞分析

细胞活力和细胞毒性测定用于药物筛选和化学物质的细胞毒性测试。Dojindo开发了高度水溶性的四唑盐,称为WST。WST-8是高度稳定的WST,用于Cell Counting Kit-8(CCK-8)。由于WST-8甲maz是水溶性的,因此不会形成晶体。因此,不需要诸如MTT测定的增溶过程。此外,CCK-8的检测灵敏度高于其他四唑盐,例如MTT,XTT,MTS或WST-1。

WST检测机制

 

ß-半乳糖苷酶检测试剂

细胞增殖/细胞毒性转染细胞染色细胞内荧光探针细菌染色微生物活力测定干细胞分化SPiDER-ßGal线粒体检测细胞代谢

应用 产品展示
细胞生长检测,药物筛选,比色/荧光检测 细胞计数试剂盒-8
细胞计数试剂盒8 + 96孔有机硅定向剂
细胞计数试剂盒-F
细胞毒性LDH检测试剂盒
-WST 96孔有机硅定向剂
MTT
了解检测机制的差异: 点击这里
细胞周期分析 细胞周期测定溶液深红色
细胞周期测定溶液蓝色