卡尔科弗卢尔荧光增白剂  Calcofluor White Stain

——酵母、真菌、寄生生物荧光染料

基本描述：

卡尔科弗卢尔荧光增白剂（Calcofluor White Stain）一种用于快速检测酵母、真菌和寄生生物的荧光染料。

作用机制（阐述）：

卡尔科弗卢尔荧光增白剂（Calcofluor White Stain）是一种与真菌和其他生物体细胞壁中的纤维素和壳多糖结合的非特异性荧光染料，卡尔科弗卢尔荧光增白剂染色是一种快速检测方法，可检测许多酵母菌和病原性真菌比如微孢子虫（Microsporidium）、棘阿米巴（Acanthamoeba）、肺囊虫（Pneumocystis）、纳氏虫（Naegleria）和阿米巴（Balamuthia）物种。

伊文斯兰（Evans blue）在该染料中用作复染剂，当使用蓝色光激发（并非UV）检测可降低组织和血细胞内的背景荧光信号。300-440nm的光谱范围（Emax=433nm0.1M磷酸盐pH 7.0；纤维素）可用作发射光谱，激发波长约在355nm。

产品组分：

卡尔科弗卢尔白M2R（Calcofluor White M2R）1 g/L

伊文斯兰（Evans blue） 0.5 g/L

染色步骤：

1. 将待检样本置于一干净载玻片上；

2. 滴一滴卡尔科弗卢尔荧光增白剂（Calcofluor White Stain）和一滴10%氢氧化钾；

3. 将盖玻片置于样本上，并静置染色1min；

4. 使用x100 to x400放大倍数紫外光下观察。

染色结果（阐述）：

真菌或寄生生物呈亮绿到蓝色荧光，而其他材料呈红-橙色荧光。若染色组织样本会有非特异性反应。对于这些样本可能会看到黄绿色背景荧光，但真菌和寄生结构荧光强度更明亮。

Fungal or parasitic organisms appear fluorescent bright green to blue, while other material is reddish-orange fluorescent. Non-specific reactions may occur when tissues samples are used. May a yellowish-green background. fluorescence is observed with such specimens but fungal and parasitic structures appears with much more intense. Attention cotton fibers will fluoresce strongly and must therefore be differentiated from fungal hyphae. As well amebic cysts are fluorescent but trophozites will not stain or fluoresce, Background fluorescence can be diminished by examining under blue light or by using different filter combinations (emission and excitation filters). One drop of 10% potassium hydroxide solution can be added for better visualization of fungal elements.

订购信息：即用型的染料，含Calcofluor White M2R和Evans blue，操作极其简单。进口原料，品质保证，现货销售。

另提供荧光增白剂Calcofluor White M2R（Fluorescent Brightener 28，CAS NO. 4404-43-7）578元/1g，冻干粉形式，更加经济实惠，欢迎来电咨询。电话：021-54736159 18121428569，QQ：2971634497。

注意事项

所有产品仅用于科研实验；

为了您的安全和健康，请穿实验服并戴一次性手套操作。

 — —Written/Edited by V. Shallan【版权归MKBio懋康所有】

上海懋康生物科技有限公司是一家涉足于生命科学和生物技术领域研究的试剂、仪器和实验室消耗品与实验服务工作，主要从事细胞生物学、植物学、分子生物学、免疫学、生物化学、蛋白组学。生物制药与诊断试剂研发生产等领域。 本公司秉承“以人为本，以诚为信、合同守信”的经营理念。坚持"品质保障"的原则为广大客户提供优质产品。

文献引用：

[1]Yu C et al. Nanoprobe-based force spectroscopy as a versatile platform for probing the mechanical adhesion of bacteria. Nanoscale. 2019 Apr 23;11(16):7648-7655. PMID: 30720812

Fluorescent Brightener 28 wasobtained from Maokang Biotechnology (Shanghai, China). For fluorescence staining of extracellularpolymeric substances (EPSs), after the biofilm formation, thePDMS surfaces were incubated with 6 µM SYTO 9 at roomtemperature in the dark for 12 min, followed by a second stain-ing of 0.15 mM fluorescent Brightener 28 for 3 min.

[2]Shengnan Tao et al. Ultra-stable Pickering emulsion stabilized by a natural particle bilayer. Chem. Commun., 2020,56, 14011-14014 https://doi.org/10.1039/D0CC05690A

Calcofluor White was bought from Maokang Biotechnology (China) Co., Ltd. Calcofluor White (SNCs) and Nile Blue A (ZNPs) were used to stain Pickering emulsions. Lasers of 405 nm and 633 nm were used to excite the dyes of Calcofluor White and Nile Blue A, respectively.

[3]Qian Li et al. 2‐Hydroxy‐4‐methoxybenzaldehyde inhibits the growth of Aspergillus flavus via damaging cell wall, cell membrane, manipulating respiration thus creating a promising antifungal effect on corn kernels. https://doi.org/10.1111/ijfs.14617

Calcofluor-White Stain (CAS: 4404-43-7) was purchased from Maokang Biotechnology Co. (Shang-hai, China).

[4]Ya Fei Geng, et al. An innovative role for luteolin as a natural quorum sensing inhibitor in Pseudomonas aeruginosa,Life Sciences,Volume 274,2021,119325,ISSN 0024-3205,https://doi.org/10.1016/j.lfs.2021.119325.

Afterwards, 50 μg/mL of Calcofluor white M2R (Maokang, Shanghai, China), a blue fluorescent marker, was used to visualize the exopolysaccharides in the biofilm matrix by staining for 2 h at room temperature in the dark.

[5]Li, Q., Zhu, X., Xie, Y. et al. o-Vanillin, a promising antifungal agent, inhibits Aspergillus flavus by disrupting the integrity of cell walls and cell membranes. Appl Microbiol Biotechnol 105, 5147–5158 (2021). https://doi.org/10.1007/s00253-021-11371-2

 Calcofluor white (CAS: 4404-43-7) was purchased from Maokang Biotechnology Co. (Shanghai, China) 

[6] JiashunShi et al. Constructing zwitterionic nanofiber film for anti-adhesion of marine corrosive microorganisms. Journal of Materials Science & Technology. Volume 70, 20 April 2021, Pages 145-155

Calcofluor stain was purchased from Maokang Biotechnology Co., Ltd (Shanghai, China). 

[7]QianLi et al. Antifungal efficacy of paeonol on Aspergillus flavus and its mode of action on cell walls and cell membranes. LWT Volume 149, September 2021, 111985

Calcofluor white (CW) (CAS: 4404-43-7) were purchased from the Maokang Biotechnology Co., Ltd 

Fig. 1. Effect of paeonol on cell walls' integrity of A. flavus. (A) SEM observation of the mycelial morphology. Quantification of the content of (B) β-1,3-glucan and (C) chitin in mycelia cell walls. a-c significant difference (P < 0.05) according to Duncan's multiple range test. (D) Distribution of septa in mycelia treated with paeonol after staining with calcofluor white under a fluorescence microscope. Scale bar: 10 μm.

https://ifst.onlinelibrary.wiley.com/doi/abs/10.1111/ijfs.14617

[8] Jing Wang et al. International Journal of Biological Macromolecules Structural and emulsion-stabilizing properties of the alkali-extracted arabinoxylans from corn and wheat brans. International Journal of Biological Macromolecules. Volume 251, 1 November 2023, 126190

Calcofluor white M2R was purchased from Maokang Bio-Technology Company (Shanghai, China).

[9] Wang J, Hou X, Hannachi K, Fan M, Li Y, Qian H, Wang L. A multi-scale approach to arabinoxylan-based emulsions: From molecular features, interfacial properties to emulsion behaviors. Int J Biol Macromol. 2024 Feb;258(Pt 1):128881. doi: 10.1016/j.ijbiomac.2023.128881. Epub 2023 Dec 21. PMID: 38134997.

 Nile red was obtained from Sigma-Aldrich Company (St. Louis, MO, America) and Calcofluor white M2R (CW M2R) was purchased from Maokang Bio-Technology Company (Shanghai, China). 

[10] Wenxiu Teng et al. Modulating the textural and structural characteristics of curdlan-potato protein gel with different concentrations. Food Hydrocolloids. Volume 149, April 2024, 109638

Rhodamine B and calcofluor white were purchased from Sigma-Aldrich, Inc. (Shanghai, China) and MaoKang Biotechnology Co., Ltd. (Shanghai, China).

[11] Du, G., Shao, Y., Luo, B. et al. Compliant Iontronic Triboelectric Gels with Phase-Locked Structure Enabled by Competitive Hydrogen Bonding. Nano-Micro Lett. 16, 170 (2024). https://doi.org/10.1007/s40820-024-01387-4

Calcofluor White Stain was purchased from Shanghai Maokang Biotechnology Co., LTD.

[12] Zhang W, He D, Wei Y, Shang S, Li D and Wang L (2022) Suppression of Aspergillus fumigatus Germination by Neutrophils Is Enhanced by Endothelial-Derived CSF3 Production. Front. Microbiol. 13:837776. doi: 10.3389/fmicb.2022.837776

Methenamine silver staining (MST, BA4094, BASO diagnostics, Inc., China) and Calcofluor white staining (CWS, 4404-43-7, Shanghai Maokang Biotechnology Co., Ltd., China) were used to label A. fumigatus.

[13] Wang Y, Fan J, Zhou Z, Goldman GH, Lu L, Zhang Y. Histone acetyltransferase Sas3 contributes to fungal development, cell wall integrity, and virulence in Aspergillus fumigatus. Appl Environ Microbiol. 2024 Apr 17;90(4):e0188523. doi: 10.1128/aem.01885-23. Epub 2024 Mar 7. PMID: 38451077; PMCID: PMC11022558.

To visualize the chitin content in the cell wall, samples were stained with 5-mg/mL CFW (Maokang Biotechnology, MS4040) in PBS for 1 min. Fluorescence microscopy was conducted after the incubation using a Zeiss Axio imager A1 microscope (Carl Zeiss, Germany) to capture the images.

[14] Qian Li, et al. Antifungal properties and mechanisms of three volatile aldehydes (octanal, nonanal and decanal) on Aspergillus flavus. Grain & Oil Science and Technology. Volume 4, Issue 3, September 2021, Pages 131-140

To investigate the influence of the three volatile aldehydes on the cell wall, Calcofluor White (CW, CAS: 4404-43-7, Maokang Biotechnology Co., Ltd., Shanghai, China) was used for assessment of any changes to cell wall integrity 

[15] Jianming Cheng et al. Application of ultrasound-assisted NaClO in membrane bioreactors membrane cleaning. Desalination and Water Treatment. Volume 264, July 2022, Pages 12-23

Concanavalin A (ConA) and Calcofluor white (CW) were purchased from Maokangbio (Shanghai, China).

[16]Liang Z, Luo Z, Zhang W, Yu K, Wang H, Geng B, Yang Q, Ni Z, Zeng C, Zheng Y, Li C, Yang S, Ma Y, Dai J. Synthetic refactor of essential genes decodes functionally constrained sequences in yeast genome. iScience. 2022 Aug 18;25(9):104982. doi: 10.1016/j.isci.2022.104982. PMID: 36093046; PMCID: PMC9460170.

Calcofluor white Cat# MM1011

[17] Dong Z, An L, Lu M, Tang M, Chen H, Huang X, Hou Y, Shen G, Zhang X, Zhang Y, Xia Q and Zhao P (2021) SPINK7 Recognizes Fungi and Initiates Hemocyte-Mediated Immune Defense Against Fungal Infections. Front. Immunol. 12:735497. doi: 10.3389/fimmu.2021.735497

 About 10 μL protein sample (5 mg mL^-1) and 10 μL heat-killed spores (1×10⁵ conidia mL^-1) labeled with Calcofluor White M2R (MKbio, China) were added to the hemocyte monolayer and incubated at 25°C for 2 h