Triplet cell division of a pancreatic tumor cell

I previously posted at

http://works.bepress.com/gmcnamara/21/

a triplet cell division of "U251t glioma cell division to triplets, acquired in 2005 by Prof. Laurence Cooper, Dr. Cludia Kowolik and George McNamara, at City of Hope National Medical Center, Duarte, CA, using CoH's Video Timelapse Light Microscopy Facility (VTLM). 3 hour video, one frame every two minutes." (that video was probably acquired with a 10x objective lens).

Prof. Cooper moved to Houston in 2006, I moved to Miami in 2007, and Cooper recruited me to Houston in 2013. Now i Sept 2014 I am doing timelapse imaging of pancreatic tumor cells -- in preparation for adding antigen specific cytolytic T-lymphocytes (CTLs, aka 'killer T-cells') to observe and quantify "serial killing". During imaging of this experiment our colleague, Dr. Ana Korngold, pointed out this triplet cell division (mid-field of view, about 50% of the way through this video). My thanks to Dr. Shan Zong, Hiroki Torikai and Amir Alpert, in our lab for this collaboration - especially Shan for providing these cells.

Due to our experiment program, I cannot provide any details of the pancreatic tumor cells used here. The entire triplet division took about 90 minutes, the normal (2 daughter) cell divisions may be faster.

Time interval: 2 minutes.

Playback: 10 frames per second.

Time duration: 1350 min (675 frames * 2 min interval), equals 22.5 hours.

Magnification 20x.

Pixel size is 500 nm (0.5 um)

Field of view 404x456 pixels, so 202x228 um (I usually use 'round number' regions of interest [ROIs], not this time).

Camera: Hamamatsu FLASH4.0 sCMOS (2,048x2048 pixel camera).

Microscope: Leica DMI6000.

Objective lens and mode: HCX PL Fluotar 20x/0.40NA and phase contrast

(I use a Leica HC Plan Apo 20x/0.75 when doing fluorescence imaging, this dataset is only transmitted light so I used the phase contrast objective lens and imaging mode).

Live cell incubator: PECON.

Imaging dish: Mattek 35 mm diameter, 20 mm diameter imaging area, #1.5 coverglass.

Culture media: DMEM with Glutamax (3 mL).

Video format: AVI file using Intel IYUV format.

I may post the entire dataset in the future (2048x2048 pixels, still 2 min interval, undecided on how long to acquire for).

George McNamara, PhD

Single Cells Analyst / Sr Research Scientist

L.J.N. Cooper lab

MD Anderson Cancer Center

Houston, TX

p.s. Dr. Ana Korngold also noticed something often seen with human cells in tissue culture: the nuclei and nucleoli sometimes look like human faces. I have posted about this previously, see

http://works.bepress.com/gmcnamara/25/

for a Halloween "trick or treat" package, where you can self-identify which cell looks like you, your friends, your ex-friends if you point out certain resemblances, your colleagues, etc (great to do during committee meetings, if you run out of faces, try self-identifying and committee-member identifying by psychogeometrics using this score card

http://www.willamette.edu/dept/careers/pdf/Psychogeometrics%20Willamette%202010.pdf

(stay on page 1 until you have identified everyone!).