human hepatoma cell line hep3b  (ATCC)

Journal: bioRxiv

Article Title: Light-dependent cell fixing with DNA-targeting fluorophores

doi: 10.64898/2026.03.27.714905

Figure Lengend Snippet: (A) Localization of PAL (green) at mtDNA within MitoRed ® -stained mitochondria (red) in live Hep3B cells prior to irradiation (scale bar: 10 µm, crop: 5 µm). (B) Top: snapshots of time-lapse PAL nuclear fluorescence upon WF irradiation (scale bar: 20 µm). Bottom: PAL labeling upon irradiation at distinct phases of the cell cycle ( a : interphase; b : prophase; c : prometaphase; d : metaphase; e : anaphase. Scale bar: 5 µm). Right : kinetics of PAL fluorogenesis during 5 min irradiation. Mean intensities are rescaled from 0 to 1 within a 95% confidence interval. (C) PAL nuclear fluorescence (in green) of a targeted cell islet within a population of live PAL-treated cells upon WF irradiation (PAL + hv) (R1: dotted line; R2: dashed line; NA: non-affected area. Scale bar: 50 µm). Crops in R1 (orange squares) showing stable nuclear morphology and persistence up to 9 days post-irradiation within a live-cell proliferative environment. Crops in R2 (blue squares) showing compromised nuclei. Calculated irradiation powers were 3.5 to 15 W/cm 2 in R1 from limit to center of the beam. Only evanescent or residual scattered light was present in R2 (scale bar: 20 µm). (D) Co-staining of PAL-treated cells (green) with propidium iodide (PI, red) or NucView ® 530 Red Caspase-3 dye (NucView, magenta). Top: PI stains R1 cells (dotted lines), showing a permeabilized status immediately post-irradiation (d0). After 24 h, R2 cells show important delayed PI staining demonstrating a compromised state. Bottom: NucView faintly stains R1 cells 24 h post-irradiation, showing strong signal in compromised R2 cells. The apoptosis inducer staurosporine is used as a positive control. (E) Quantification of nuclear size in different conditions: live PAL-treated cells without irradiation (PAL), FA-fixed cells (FA), PAL-free irradiated cells (hv) or PAL-treated irradiated cells (PAL + hv, R1 and R2) (n>200 per condition). Data shown in violin plots reflecting size distribution, circles indicate the medians and whiskers indicate standard deviations.

Article Snippet: The human cell line Hep3B was obtained from ATCC ® (HB-8064), the human cell lines T24, PC3 and C4-2B were a courtesy of Pr.

Techniques: Staining, Irradiation, Fluorescence, Labeling, Positive Control

Journal: bioRxiv

Article Title: Light-dependent cell fixing with DNA-targeting fluorophores

doi: 10.64898/2026.03.27.714905

Figure Lengend Snippet: (A) Left: Optofixing procedure of a whole Hep3B cell population using a portable LED lamp towards biochemical analysis. Top right: Visualization of abundance of precipitates (UVA irradiation). Right bottom: Corresponding SDS-PAGE analysis of soluble protein contents following treatments with formaldehyde (FA), 4-hydroxynonenal (4HNE), acrolein (ACR) or LED irradiation in presence or absence of PAL. (B) Left panel: WF imaging of PAL nuclear fluorogenesis at day 0 after irradiation by a WF microscope. Right panel: stability of fixed state after 3 days of PAL-treated cells in presence of inhibitors (scale bar: 10µm). (C) Left: fluorogenesis kinetics during 5 min irradiation with conditions described in B . Fluorescence intensities were normalized as [(I-Io)/Io]. Right : quantification of nuclear sizes with conditions described in B (n>60 per condition) 3 days post-irradiation. ( D) Lipid peroxidation (LPO)-quantification in PAL-treated cells using BODIPY™ 581/591-C11 probe. Top left: representative image of probe staining 30 min after irradiation of PAL-treated cells in zones NA, R1 and R2. Top right: plot of ratiometric measurements along the yellow dashed line. Red curve represents Red/Ox ratio, green curve represents the Ox/Red ratio. Bottom: quantification of the oxidation ratio (Ox/Red) 30 min after irradiation of PAL-treated cells (R1, R2) compared to irradiated cells in absence of PAL (hv) or cells treated with PAL without irradiation (PAL) (n>100 per condition). (E) ROS quantification in PAL-treated cells using ROS Brite TM 670 probe. Top: representative image of ROS staining (in magenta) obtained after irradiation of PAL-treated cells (in green) in R1 and R2 (scale bar: 10 µm). Bottom: Quantification of ROS Brite fluorescence intensity in the same conditions than D .

Article Snippet: The human cell line Hep3B was obtained from ATCC ® (HB-8064), the human cell lines T24, PC3 and C4-2B were a courtesy of Pr.

Techniques: Irradiation, SDS Page, Imaging, Microscopy, Fluorescence, Staining

Journal: bioRxiv

Article Title: Light-dependent cell fixing with DNA-targeting fluorophores

doi: 10.64898/2026.03.27.714905

Figure Lengend Snippet:

Article Snippet: The human cell line Hep3B was obtained from ATCC ® (HB-8064), the human cell lines T24, PC3 and C4-2B were a courtesy of Pr.

Techniques: Labeling