Gleiberman, Anatoli (2021). CIL:814, Homo sapiens, permanent cell line cell. In Cell Image Library. UC San Diego Library Digital Collections. Dataset. https://doi.org/10.6075/J0GX49SN

This culture of HeLa cells was grown, fixed and labeled directly on 35-mm plastic petri dishes (not on cover slips). To preserve the 3D structure of cells, cultures were fixed in formaldehyde vapors at room temperature over 20 min. Fixation was performed as follows: medium was completely aspirated, a 50ul drop of 37% formaldehyde was placed on the lid of the dish and covered with the dish of cells, bottom up. After fixation dishes were carefully filled with 1ml of 50% glycerol in PBS and dishes were stored at -20 C until staining.

Before staining, cells were incubated with blocking solution (5% fetal calf serum, PBS, 0.2% triton x-100, 25 min room temp.), then treated with primary antibody, which was mouse anti-Cytochrome C from BD Pharmigen (cat.# 556433) diluted 1:100 in blocking solution, 1h room temp. After standard washing (PBS 3-4 times, 5 min each), secondary antibody (donkey monovalent Fab-fragment anti-mouse IgG conjugated with Rhodamine Red-X from Jackson ImmunoResearch laboratories, concentration 2.5ug/ml) was applied.

Images were captured on a Zeiss Axioplan-2. Objective - 63x oil immersion/NA 1.25. Magnifying lens (optovar) was 1.5. Digital camera – Hamamatsu ORCA-ER. Program – AxioVision 3.5.1. Illumination-mercury bulb HBP50. Filter set – standard for Rhodamine/Cy3/Alexa 564 – excitation 530-585 and emission LP 615. Mounting medium-VectaShield.

• 2021

• Gleiberman, Anatoli

Preparation: formaldehyde fixed tissue; glycerol permeabilized; detergent permeabilized; primary antibody plus labeled secondary antibody
Relation to intact cell: whole mounted tissue
Item type: microscopy
Imaging mode: fluorescence microscopy
Parameter imaged: fluorescence emission
Source of contrast: local accumulation of fluorescent probe
Visualization methods: Rhodamine
Processing history: unprocessed raw data

• Homo sapiens

• Mitochondrion
• Permanent cell line cell

• Apoptotic mitochondrial changes
• ATP biosynthetic process

• data
• image

• No linguistic content; Not applicable

Samplenumber: 814

Doi: https://doi.org/10.6075/J0GX49SN

Source data

• Source Record in the Cell Image Library: https://doi.org/10.7295/W9CIL814

CC0 1.0 Universal Public Domain Dedication

Creative Commons Public Domain Dedication

Use: The person(s) who associated a work with this deed has dedicated the work to the public domain by waiving all of their rights to the work worldwide under copyright law, including all related and neighboring rights, to the extent allowed by law.

Constraint(s) on Use: This work may be used without prior permission.

Research Data Curation Program, UC San Diego, La Jolla, 92093-0175 (https://lib.ucsd.edu/rdcp)

2022-08-12