Cell lines in 3D

PANC-1

On the photos below there is results of Panc-1 triple staining designed for staining different cellular organels – DiO (green) for cell membranes, MitoLite (red) for mitochondria and Hoechst (blue) for nuclei.

MiaPaca-2

Cells from photo on the left and in the middle have been cultured on LifeGel 48-well plates for 10-14 days, 2500 cells per well have been seeded. On the photo on the right, cells were cultured for 10 days on 96-well plate, they have been seeded 1000 cells per well.

CFPAC-1

CFPAC-1 cells have been cultured on 96-well plate with LifeGel for 14 days.

Pan_02 cells 3D structures immunofluorescence staining of actin (green), tubulin (red) and nuclei (blue) growing on LifeGel.

Cells were fixed using 4% formaldehyde solution (incubation 10-30 minutes), then perforated with 0,1% Triton-X in 10% FBS (in PBS). Actin was stained using DY-490-Phalloidin from Dynomics (1:500 in PBS). Tubulin was stained using primary (Monoclonal anti-alpha-Tubulin antibody, mouse; 1:500) and secondary (Anti-mouse IgG-Atto594; 5ug/mL) antibodies. Nuclei were stained with Hoechst 33342 (10 ug/mL). Photos: obj. 10x lens 1x.

RKO cells have been cultured on 96-well LifeGel plates for 12 days, they have been seeded 1500 cells/well.

4T1 cells shown below have been cultured on µ-Slide angiogenesis plate with LifeGel for 7 days. Cells were seeded 500 cells/well.

On the photos below there is staining with cultured 4T1 cells on 96-well LifeGel plate. DiO was for cell membranes (green), MitoLite for mitochondria (red), Hoechst for nuclei (blue). Obj. 2x, lens 1x.

Cells have been fixed on LifeGel and fluorescently stained with antibodies for: F-actin (green colour), α-tubulin (red colour), nucleus (blue colour). Photos in the second line is with brightfield and darkfield.

Project have been discovered in cooperation with IBIDI. Photos made by Shokoufeh Teymouri, µ-Slide Angiogenesis, IBIDI GmbH.

Photos below show U-2 OS cells with stable transfection of GFP growing on LifeGel. Cells have been cultured on 96‑well plate for 21 days, 500 cells per well was seeded (obj. 2,5x lens 1,6x).