HS-CyBz

Operating Instructions (The following is a recommended experimental protocol for guidance only, and adjustments are required based on specific requirements)

1. Stock Solution Preparation

2. Working Solution Preparation
2.1 Working concentration: 20 μM.
2.2 Notes: Adjust the working solution concentration as needed; prepare fresh immediately before use.

3. Staining Procedure
3.1 Sample types: Live mouse tissues; ex vivo mouse liver tissues^[1].
3.2 Incubation conditions:
3.2.1 For mouse subcutaneous tissue imaging: Use 20 μM probe solution, and incubate for 30 min after analyte injection.
3.2.2 For in vivo/ex vivo mouse liver imaging: Inject the probe intravenously 12 h after intraperitoneal injection of SAM.
3.3 Washing step: No washing step is specified.

4. Control Setup
4.2 Set up negative controls, positive controls and blank controls.
4.2.1 Negative control: Inject normal saline into parallel mouse tissue regions instead of Na₂S or SAM.
4.3 Positive controls are used to validate the experimental system; blank controls are used to eliminate fluorescent interference from reagents.

5. Detection and Analysis
5.1 Instruments:
5.1.1 Fluorescence imaging: Ex 560 nm, Em 620 nm and 790 nm; Ex 720 nm, Em 790 nm.
5.1.2 Photoacoustic imaging: Ex 775 nm and 825 nm.
5.2 Result analysis:
5.2.1 Fluorescence intensity changes: In response to H₂S, the fluorescence intensity at 620 nm increases while that at 790 nm decreases, leading to an increased F₆₂₀/F₇₉₀ ratio.
5.2.2 Photoacoustic signal changes: In response to H₂S, the PA signal at 775 nm decreases, leading to an increased PA₈₂₅/PA₇₇₅ ratio.
5.2.3 Fluorescence localization: After intravenous injection, the probe accumulates in mouse liver tissues.
5.2.4 Expected ratio changes: SAM induces upregulation of endogenous H₂S in mouse liver, resulting in higher F₆₂₀/F₇₉₀ and PA₈₂₅/PA₇₇₅ ratios compared with the normal saline-treated control group.

MedChemExpress (MCE) has not independently confirmed the accuracy of these methods. They are for reference only.