Labeling cells, organelles, toners, viruses, and lipoproteins with henna dyes is a common practice. The dialkylphoenix dye DiA (4-Di-16-ASP), which is used to designate membranes and other focal structures, is one of the long-chain henna cyanines, along with DiO (DiOC18 (3)), DiI (DiIC18 (3)), DiD (DiIC18 (5)), and DiR. Comparing DiIC16 (3) to DiI (C18), the alkyl substituent (C16) is shorter. They exhibit brief excited-state depletion in pyramid settings, environment-dependent fluorophores, and extraordinarily high extinction factors. While they are oil-forming and have poor fluorescence in water, when they are attached to lipophilic biomolecules or integrated into membranes, they become moderately photostable and highly fluorescent. Their optical characteristics render them perfect for the staining of cell plasma membranes. These dyes diffuse laterally within the plasma membrane after cells are identified, staining the entire cell [1 DiO, DiI, DiD, and DiR exhibit distinct green, orange, red, and red fluorescence, respectively, making it easier to image live cells in multiple colors and perform flow cytometry analysis. You can utilize normal FITC and TRITC filters with DiO and DiI, respectively. Because they often show extremely low cytotoxicity, DiI and its analogs are the most widely employed among them. Furthermore, lipoproteins like LDL and HDL are frequently measured using DiI. Another often used tool for determination is the lipophilic radical dye DiA. Protocol general 1. Get Di staining solution ready. The Metatracer [2]. Compile stock solutions in dimethylformamide (DMF), dimethylformamide (DMSO), or ethanol DMSO. The concentration of the stock solutions should range from 1 to 5 mM. To act as Di's solvent, DMF b. Aliquot the stock solution and store it in the freezer at a minimum of -20°C. An aliquot of any leftover solution is advised. Don't freeze and thaw again. The solution has a six-month shelf life. B. To create a working solution, mix the stock solution with a suitable buffer, like serum-free medium, HBSS, or PBS, ranging from 1 to 5 μM. Storing the sink for longer than a day is not advised. Note: Two is the working solution's final concentration. Cells suspended a. For three to five minutes, centrifuge at 1000 g at 4°C. Once the Di working solution has been added, wait one to thirty minutes. D. Rinse twice with PBS for five minutes each time, then centrifuge three to four times at 400 g and 4°C. Discard the supernatant. D. Wash for five minutes each time, twice, using PBS. E. After resuspending the cells in PBS or serum-free cells, examine them using a microscope or flow cytometry. 3. cells that adhere a. B. After removing the coverslip from the culture medium, aspirate any extra cells. an. Once the cells are well covered, add 100 μL of the working solution, shake gently, and let the cells purify for five to thirty minutes. Apply culture medium twice, allowing each wash to last five minutes. Observe using flow cytometry or fluorescence microscopy.

[1]. Multicolor "DiOlistic" labeling of the nervous system using lipophilic dye combinations. Neuron. 2000 Aug;27(2):219-25.

[2]. Applied electric fields accelerate the diffusion rate and increase the diffusion distance of DiI in fixed tissue. J Neurosci Methods. 2005 Jan 30;141(1):155-63.

DiIC18(7) dye is an organic iodide salt. It has a role as a fluorochrome. It contains a diIC18(7)(1+). It is functionally related to a C7-indocyanine.

C63H101IN2

1013.3939

1012.7

100068-60-8

25195411

Brown to reddish brown solid powder

0

2

38

66

1260

0

CCCCCCCCCCCCCCCCCCN\1C2=CC=CC=C2C(/C1=C\C=C\C=C\C=C\C3=[N+](C4=CC=CC=C4C3(C)C)CCCCCCCCCCCCCCCCCC)(C)C.[I-]

JLIOTPLALDYAEH-UHFFFAOYSA-M

InChI=1S/C63H101N2.HI/c1-7-9-11-13-15-17-19-21-23-25-27-29-31-33-38-46-54-64-58-50-44-42-48-56(58)62(3,4)60(64)52-40-36-35-37-41-53-61-63(5,6)57-49-43-45-51-59(57)65(61)55-47-39-34-32-30-28-26-24-22-20-18-16-14-12-10-8-2;/h35-37,40-45,48-53H,7-34,38-39,46-47,54-55H2,1-6H3;1H/q+1;/p-1

(2E)-2-[(2E,4E,6E)-7-(3,3-dimethyl-1-octadecylindol-1-ium-2-yl)hepta-2,4,6-trienylidene]-3,3-dimethyl-1-octadecylindole;iodide

2934.99.9001

Powder      -20°C    3 years

                     4°C     2 years

In solvent   -80°C    6 months

                  -20°C    1 month

Note: Please store this product in a sealed and protected environment (e.g. under nitrogen), avoid exposure to moisture and light.

Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs)

DMSO : ~25 mg/mL (~24.67 mM)

Solubility in Formulation 1: ≥ 2.08 mg/mL (2.05 mM) (saturation unknown) in 10% DMSO + 90% (20% SBE-β-CD in Saline) (add these co-solvents sequentially from left to right, and one by one), clear solution.
For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 20.8 mg/mL clear DMSO stock solution to 900 μL of 20% SBE-β-CD physiological saline solution and mix evenly.
Preparation of 20% SBE-β-CD in Saline (4°C，1 week): Dissolve 2 g SBE-β-CD in 10 mL saline to obtain a clear solution.

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 (Please use freshly prepared in vivo formulations for optimal results.)