2022 Eureka

2022 Eureka kdorfman Thu, 07/14/2022 - 19:27

color mixing workshop LLCPK GFP-alpha tubulin cells (have a lot of vacuoles, though)

Made up 2x staining solutions in Fluorbrite:

Nuc blue: 20 uL/2 mL; 40 uL/2 mL

Mitotracker red: 250 nM, 500 nM

Cells incubated in coverslip dish 1 mL nuc blue staining solution + 1 mL mitotracker staining solution.

Hi nuc + Hi MT
Hi nuc + low MT
low nuc + Hi MT
low nuc + low MT

Warm all media first, then remove the F10-Hams, replace with appropriate staining solutions. Keep cells in incubator for 15 min, check...

Color Mixing with Inks CMY(K)

1:15 (approximate) yellow printer ink in water 1:200 magenta 1:200 cyan

we pipetted various ink amounts onto parafilm to mix colors - final volume of droplets = 100ul

Additive Mixing with Light (RGB Flashlights) Flashlights made by RaySoar LED Limited Company

Staining solution NucBlue + Mitotracker

Staining solution NucBlue + Mitotracker kdorfman Fri, 09/30/2022 - 13:47

Staining Solution

  • 1 mL fluorobrite
  • 10uL NucBlue
  • 1 uL Mitotracker red

Staining Protocol

  • Remove pink medium from cells (~2.5 mL)
  • Replace with Staining Solution
  • Rock dish to make sure solution covers the surface
  • Incubate ~15 min at 37C

Observation Protocol

  • Find cells with phase
  • Switch to fluorescence
    • turn on arc lamp
    • turn off white light
    • open mechanical shutter (CLOSE WHEN NOT LOOKING AT CELLS! THEY BLEACH!)
    • open electronic shutter
    • turn filter cube to UV to see blue nuclei
    • turn filter cube to G to see red mitochondria