FlashFix - Optofixation solution
Solution for cell fixation with high spatial and temporal resolution.
A technology originally developed by Stephanie Bosch (1), Maelle Carraz (1,2), Romain Duval (2)
(1) Centre de Biologie Intégrative (CBI), Unité de Biologie Moléculaire, Cellulaire et du Développement (MCD), CNRS, Toulouse, France
(2) Institut de Recherche pour le Développement (IRD), Paris , France
FlashFix, a photofixative technology.
FlashFix is a new way of fixing cell with high spatial and temporal resolution. It is based on a molecule which can trigger cell fixation when exposed to light in the proper culture conditions.
Fast fixation
Cells are snap fixed, down to 7 seconds
Precision
Resolution of fixation down to single cell
Visible
Nuclei are stained in green after photofixation
How does FlashFix work ?
Applications
- Snap fix the cells at a precise point to capture rare cellular events.
- Study the behaviour of surrounding cells when a group of cells are fixed, similarly to photo-ablation
Validated samples
FlashFix has been successfully used on a variety of cells :
- Hep3B : liver cancer cell line
- C4-2B : prostate cancer cell line
- PC3 : bone metastasis cell line of a grade IV prostatic adenocarcinoma
- T24 : urinary bladder cell line of colorectal adenocarcinoma
- WI-38 : fibroblast-like fetal lung cell.
- Cells dissociated from zebrafish embryo
Kit description:
- 1000 X stock photofixing solution
- Specific cell culture media
Find out all results linked to FlashFix in the publication section !
Long-term immobilisation of cells following optofixation with FlashFix
Hep3B cells were imaged right after optofixation (day 9) and subsequently followed for up to 9 days. Nuclei of fixed cells are represented in green. The absence of movement of nuclei after time shows the efficiency of fixation process. In another set of experiments, a reduction of lipid droplet movement also indicated a global absence of any intracellular movement. Scale bar = 20µm.
High control over fixation spatial resolution with FlashFix
Upon illumination with 488nm light, cells were locally fixed with FlashFix. The left panel shows Hep3B cells visible in bright field. The middle and left panels show the GFP signal of cells incubated with FlashFix before (middle panel) and after (right panel) irradiation. Cells centered within the irradiation beam were strongly labeled in green, which is an indicator of their fixed status. Scale bar = 50µm.
Fixation with FlashFix is correlated with nuclear fluorescence
FlashFix fixative compound becomes fluorescent when it binds to DNA. Under normal culture conditions, it can partially diffuse into mitochondria and stains very faintly mitochondrial DNA. After light irradiation, the cells become permeabilized, allowing the FlashFix compound to enter the nuclei, which then emit green fluorescence. The appearance of nuclear fluorescence confirms that fixation has occurred.
Capture of precise mitotic events with FlashFix
Irradiation of cells incubated with FlashFix at distinct phases of the cell cycle (Interphase, prophase, prometaphase, metaphase). Scale bar: 5 μm
Compatibility of FlashFix with immunofluorescence labeling
Comparison of immunofluorescence labeling following FlashFix and formaldehyde (FA) fixation. Representative image of nuclear membrane (LaminB1) staining. The marker is detectable in a similar manner in both conditions. Additional nuclear markers were successfully used, including euchromatin (H3Ac), heterochromatin (SUZ12), interspace chromatin (PML bodies), telomers (TRF1). Scale bar = 10µm
Versatility of FlashFix with multiple cell types
When cells are incubated with FlashFix, they will become globally fluorescent. The pattern of fluorescence can change according to the cell type and condition. Upon illumination with a 488nm laser, the fluorescence will be re-located only to the nucleus, confirming fixation. (T24 : urinary bladder of a male with colorectal adenocarcinoma; PC3 : cell line bone metastasis of a grade IV prostatic adenocarcinoma; eZF : cells dissociated from 48 hpf 5 zebrafish embryo; C4-2B : cell line prostate cancer).