Dansyl-morpholine

Guide (The following is our recommended protocol. This protocol is for guidance only and should be modified according to your specific needs).

1. Stock Solution Preparation
(No solvent or concentration information provided in literature; items omitted)

2. Working Solution Preparation
2.1 Diluent: Cell culture medium.
2.2 Working concentration: 10 μM.
2.3 Note: Adjust working solution concentration as needed; prepare fresh before use.

3. Staining Procedure
3.1 Sample type:[1]
3.1.1 Adherent cells: Seed cells on 35-mm glass-bottom dishes and culture for 24 h before staining.
3.2 Incubation conditions:
3.2.1 Living cells: Incubate with 10 μM DNS-M for 10 min; bright fluorescence is observable within 1 min.
3.2.2 Fixed cells: Fix cells with 4% neutral buffered formalin for 30 min, then incubate with 10 μM DNS-M for 10 min.
3.3 Washing steps:
3.3.1 Fixed cells: Wash with PBS three times after fixation and before probe incubation.

4. Controls
(No controls explicitly mentioned in literature; section omitted)

5. Detection & Analysis
5.1 Instrument: Laser scanning confocal microscope.
5.1.1 Excitation (Ex): 405 nm; Emission (Em): 480−540 nm.
5.2 Result analysis:
5.2.1 Fluorescence intensity changes: Cancer cells exhibit significantly brighter green fluorescence than normal cells.
5.2.2 Fluorescence localization: Green fluorescence specifically localizes to lipid droplets (LDs), with a co-localization coefficient of 0.97 when compared to Nile red.
5.2.3 Fluorescence dynamics: In cancer cells, LDs exhibit dynamic behaviors including dissociation, re-association, and fusion within minutes; in normal cells, LDs show minimal movement.
5.2.4 LDs characteristics: Cancer cells have larger and more numerous LDs than normal cells.