Wednesday, July 17, 2013
Video Introducing the 4D Technology BioCam Phase Imaging Microscope
This video shows a beta version of the BioCam Quantitative Phase Microscope in action. Our thanks to Howard Letovsky for assembling this video.
Dynamic Cellular Measurements
In a recent case, cardiac myocytes from 1-2 day old neonatal Sprague Dawley (Harlan, Indianapolis, IN) rats were prepared at the University of Arizona and plated onto #1 round coverslips. After 2 weeks of incubation the cells were measured in a Bioptechs FCS3 perfusion chamber with an aluminized base plate and a 100μm gasket. Cells were kept at 37°C with an HBSS (Hank’s balanced salt solution) fluid bath. Figure 3 shows OT maps of an area of cells imaged at 40X (20X with 2X FOV lens) before and after pushing IPHC (isoproterenol hydrochloride – a beta adrenergic agonist).
The images below are the first in two time series of 200 datasets taken at 15 fps over a 13.3 sec time period. These images are the same 349 x 326 pixel subarea of 1050x1200 images. The optical volume of the subareas is determined by summing all the OT values and then scaling so that both traces have the same mean and same relative scaling. The actual physical volume is not obtainable from these data because the thickness of the cell culture and the index of refraction data are unknown.
A major advantage of this technique is that relative physical changes can easily be measured. The figure below shows the relative optical volume for these two time series before and after pushing IPHC. These cells spontaneously beat (or twitch) about once every 4 seconds when in the HBSS at 37°C. After the IPHC is pushed, the beating frequency increases by a factor of 8 to twice a second and the flexion is about 3 times stronger. The optical volume is getting smaller in this subarea when the cells beat because the cells are noticeably stretching out and expanding during the beats.
These measurements show how quantitative optical phase and optical volume measurements can be
used to study a group of cells over time.
The images below are the first in two time series of 200 datasets taken at 15 fps over a 13.3 sec time period. These images are the same 349 x 326 pixel subarea of 1050x1200 images. The optical volume of the subareas is determined by summing all the OT values and then scaling so that both traces have the same mean and same relative scaling. The actual physical volume is not obtainable from these data because the thickness of the cell culture and the index of refraction data are unknown.
A major advantage of this technique is that relative physical changes can easily be measured. The figure below shows the relative optical volume for these two time series before and after pushing IPHC. These cells spontaneously beat (or twitch) about once every 4 seconds when in the HBSS at 37°C. After the IPHC is pushed, the beating frequency increases by a factor of 8 to twice a second and the flexion is about 3 times stronger. The optical volume is getting smaller in this subarea when the cells beat because the cells are noticeably stretching out and expanding during the beats.
These measurements show how quantitative optical phase and optical volume measurements can be
used to study a group of cells over time.
Swimming Paramecium Video
This video shows the cilia of a paramecium moving measured using the 4D Technology BioCam.
The 3D surface topographic maps which comprise the video were obtained at 15fps and 50X magnification. Cilia motion is quantifiable from time series image sequences.
The image strip below shows the paramecium using various BioCam Quantitative Phase Microscope imaging modes: (a) Phase contrast. (b) DIC. (c) Dark field. (d) OT. (e) OT/DIC Composite. (f) 3D OT topography.
Protozoa Video
This movie shows a swimming protozoa in water measured with the 4D Technology BioCam Quantitative Phase Microscope. The video shows various imaging modes, which can be changed on the fly either before or after data capture.
Rat Cardiac Myocytes
This video shows the changes in optical thickness (OT) of spontaneously beating rat cardiac myocytes as imaged with the BioCam Quantitative Phase Microscope. The myocytes were measured in a Bioptechs FCS3 perfusion chamber with an aluminized base plate and a 100μm gasket. Cells were kept at 37°C with an HBSS (Hank’s balanced salt solution) fluid bath. The video on the left shows a 2D representation of the OT data, while the video at right shows the data in 3D. The maps were acquired at 15fps.
Tuesday, July 9, 2013
Successful survey prompts product release plans
Thanks to everyone who participated in the "New Directions in Live Cell & Tissue Imaging" survey at microscopyeducation.com.
Congratulations to Scott Russell of the Noble Microscopy Lab at the
University of Oklahoma, who won our drawing for the the Bose Wave III
Radio and iPod docking station!
Over 340 of you provided valuable feedback regarding this intriguing new technology. Many of you also expressed interest in becoming beta sites for the technology.
Due to the overwhelmingly positive feedback from the survey 4D Technology is proceeding with the commercialization of this technology. 4D has a long history of creating successful products based on novel interferometric technologies. The new BioCam Quantitative Phase Microscope will extend the usefulness of these techniques into bioscience applications.
As you learned from the survey, our phase imaging technology measures optical topography along with variations in optical thickness of living cells and tissues. The BioCam will allow researchers to rapidly map and quantify optical thickness, acquiring dimensional and volumetric information at a rate of 15 frames/second, without perturbing living cells or requiring fixed material. This data provides valuable information for cellular dynamics, motility, and cell and tissue morphology.
If you have an application that you would like to test with this new technology, please let us know. We are also looking for development partners who would like to help us further our work in specific application areas and who want to explore the uniqueness of this new technology.
If you are interested in more information on the BioCam Quantitative Phase Microscopeor interested in becoming a beta site, or if you know someone who would be interested in this technology, please contact Kathy Creath, Principal Investigator for the project, at kathy.creath@4dtechnology.com or (520) 294-5600 x211. You can also connect with Dr. Creath on LinkedIn.
Over 340 of you provided valuable feedback regarding this intriguing new technology. Many of you also expressed interest in becoming beta sites for the technology.
Due to the overwhelmingly positive feedback from the survey 4D Technology is proceeding with the commercialization of this technology. 4D has a long history of creating successful products based on novel interferometric technologies. The new BioCam Quantitative Phase Microscope will extend the usefulness of these techniques into bioscience applications.
As you learned from the survey, our phase imaging technology measures optical topography along with variations in optical thickness of living cells and tissues. The BioCam will allow researchers to rapidly map and quantify optical thickness, acquiring dimensional and volumetric information at a rate of 15 frames/second, without perturbing living cells or requiring fixed material. This data provides valuable information for cellular dynamics, motility, and cell and tissue morphology.
BioCam Phase Imaging Microscope
If you have an application that you would like to test with this new technology, please let us know. We are also looking for development partners who would like to help us further our work in specific application areas and who want to explore the uniqueness of this new technology.
If you are interested in more information on the BioCam Quantitative Phase Microscopeor interested in becoming a beta site, or if you know someone who would be interested in this technology, please contact Kathy Creath, Principal Investigator for the project, at kathy.creath@4dtechnology.com or (520) 294-5600 x211. You can also connect with Dr. Creath on LinkedIn.
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