HOE-S 785026 trihydrochloride is a live cell labeling dye that binds to nucleic acids by targeting the minor groove in DNA double strands, with a preference for A/T-rich regions (fluorescence is significantly enhanced by A/T-rich double-stranded DNA). It crosses cell membranes and binds to living or immobilized cells, with its fluorescence intensity increasing as the solution pH rises.

Instructions
I. Solution preparation
1. Preparation of mother solution: Dissolve HOE-S 785026 trihydrochloride in anhydrous DMSO to prepare a 1 mg/mL mother solution. To ensure complete dissolution, the mother solution can be warmed, but high temperature should be avoided to prevent degradation.
Note: When storing the mother solution, be sure to seal it and store it at -20°C away from light.
2. Preparation of working solution: Dilute the mother solution to the required concentration in an appropriate proportion, usually using a working concentration of 5-10 μg/mL. Use serum-free cell culture medium, PBS or other suitable buffer for dilution. The specific concentration can be adjusted according to experimental needs.
II. Operation steps
1. Cell staining:
Adherent cells: First culture the cells in a culture dish to an appropriate density, add the working solution, and incubate at room temperature for 15-20 minutes.
Suspended cells: Collect the cells by centrifugation, remove the culture medium, add the working solution, and incubate at room temperature for 10-15 minutes.
During the incubation process, ensure that the cells are evenly exposed to the dye solution.
2. Washing steps:
1) After incubation, wash the cells three times with PBS or culture medium, gently centrifuging each time to remove excess dye.
2) Resuspend the cells with fresh culture medium and prepare for fluorescence detection.
3. Fluorescence observation: Use a fluorescence microscope at a wavelength of 350-370 nm for excitation and an emission wavelength of approximately 460-470 nm to observe the staining results.