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Quantitative optical microscopy - I also encourage this article

http://www.jove.com/video/50988/quantitative-optical-microscopy-measurement-cellular-biophysical
Video requires (institutional) subscription - the .M files are open access
PDF version is open access at
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4162510/pdf/jove-86-50988.pdf

abstract

We describe the use of a standard optical microscope to 
perform*quantitative*measurements of mass, volume, and density on 
cellular specimens through a combination of bright field and 
differential interference contrast imagery. Two primary approaches are 
presented: noninterferometric*quantitative**phase*microscopy (NIQPM), to 
perform measurements of total cell mass and subcellular density 
distribution, and Hilbert transform differential interference contrast 
microscopy (HTDIC) to determine volume. NIQPM is based on a simplified 
model of wave propagation, termed the paraxial approximation, with three 
underlying assumptions: low numerical aperture (NA) illumination, weak 
scattering, and weak absorption of light by the specimen. Fortunately, 
unstained cellular specimens satisfy these assumptions and low NA 
illumination is easily achieved on commercial microscopes. HTDIC is used 
to obtain volumetric information from through-focus DIC imagery under 
high NA illumination conditions. High NA illumination enables enhanced 
sectioning of the specimen along the optical axis. Hilbert transform 
processing on the DIC image stacks greatly enhances edge detection 
algorithms for localization of the specimen borders in three dimensions 
by separating the gray values of the specimen intensity from those of 
the background. The primary advantages of NIQPM and HTDIC lay in their 
technological accessibility using "off-the-shelf" microscopes. There are 
two basic limitations of these methods: slow z-stack acquisition time on 
commercial scopes currently abrogates the investigation of phenomena 
faster than 1 frame/minute, and secondly, diffraction effects restrict 
the utility of NIQPM and HTDIC to objects from 0.2 up to 10 (NIQPM) and 
20 (HTDIC) μm in diameter, respectively. Hence, the specimen and its 
associated time dynamics of interest must meet certain size and temporal 
constraints to enable the use of these methods. Excitingly, most fixed 
cellular specimens are readily investigated with these methods. PMID: 
24747818


Also IATIA (now Ultima Capital)
http://www.ultimacapital.net/iatiaimaging/Publications/Iatia%20Imaging/applicationNotes/comparisonWithOpticalPhaseContrastModalities.pdf

Yet another company, Ovizio
http://www.ovizio.com/
looks especially useful for cell bioreactors (i-Line), does have a 
camera for a microscope.

and a classic (but not trivial to implement interferometry on a research 
microscope),

Brown AF, Dunn GA 1989 Microinterferometry of the movement of dry matter 
in fibroblasts. J Cell Sci 92: 379-389.
We describe the use of interferometric microscopy coupled with a novel 
application of Sénarmont compensation for detecting and quantifying the 
distribution of*dry*matter in cultured cells. In conjunction with video 
techniques and digital image processing, a two-dimensional, calibrated 
map of the*dry**mass*distribution in an isolated cell can be obtained 
and digitally recorded. We have called the technique Digitally Recorded 
Interferometric Microscopy with Analyser Shift (DRIMAS). The method 
greatly facilitates the automatic recognition of cells by computer. 
Recorded time-lapse sequences can be used to establish a database of the 
growth and motility of specific cells in given experimental conditions. 
Databases of this type can be analysed to reveal the patterns of growth 
and locomotory behaviour of individual cells. We describe a systematic 
method of obtaining parameters of cell size, shape, spreading, 
intracellular motility and translocation. Auto-correlations and 
cross-correlations between these parameters can be detected and 
quantified using time series analysis, revealing potential cause/effect 
relationships in the mechanisms of growth and motility. Besides 
characterizing the overall pattern of cell behaviour, these data can 
also yield information about the instantaneous pattern of intracellular 
motility. We describe the use of finite element analysis to reveal the 
dynamics of the intracellular transport of*dry*matter. This yields the 
pattern of the minimum flow of*dry*matter required to account for the 
changes in its distribution. Most of this flux is not associated with 
the movement of visible structures and possibly represents the transport 
of dissociated components of the cytoskeleton. In chick heart 
fibroblasts, surprisingly high velocities of nearly 2.0 microns s-1 were 
detected during the period of increased motility following tail 
detachment. The total kinetic energy associated with the*dry**mass*flux 
is a single parameter, which characterizes the instantaneous motility of 
the cell. We found that the kinetic energy of intracellular motility can 
be several hundred times greater than the kinetic energy of 
translocation. Kinetic energy may prove to be a very informative single 
measure of intracellular motility for assessing the effects of malignant 
transformation, genetic manipulations, and other experimental treatments 
on the locomotory machinery of the cell.


On 4/15/2016 12:50 PM, Joshua Zachary Rappoport wrote:
> *****
> To join, leave or search the confocal microscopy listserv, go to:
> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
> Post images on http://www.imgur.com and include the link in your posting.
> *****
>
> Sorry everyone!
>
> Here is the link: http://view6.workcast.net/register?cpak=7302640263452844
>
> It's time for lunch...
>
> From: Joshua Zachary Rappoport
> Sent: Friday, April 15, 2016 12:06 PM
> To: [log in to unmask]
> Subject: Upcoming webinar
>
> Dear Listserv:
>
> I would like to draw your attention to a live webinar that I will be moderating starting at 9:00 am CDT on Wednesday April 27th on the subject of label-free imaging.  The webinar is being hosted by the International Journal of Biochemistry and Cell Biology (IJBCB), and sponsored by PhaseFocus.  The speakers will include Martin Humphry from PhaseFocus, Davide Danovi from Kings College London, and Peter O'Toole from the University of York, and it will be followed by a live Q&A.
>
> Label-free imaging represents an under-utilized cost-effective and user-friendly approach that has already made significant impact on fields such as cell adhesion, cell migration, cell proliferation, apoptosis and nanomaterial characterization.  Several companies offering different types of quantitative label-free imaging platforms have recently emerged including PhaseFocus, Phasics, NanoLive, PhiOptics and CytoViva.  These different solutions hold the potential to revolutionize the way we image cells, tissue and materials and I invite you all to tune into the webinar!
>
> Thanks
>
> Josh
>
>
> Joshua Z. Rappoport PhD
>
> Director of the Center for Advanced Microscopy and the Nikon Imaging Center at Northwestern University
> Northwestern University Feinberg School of Medicine
> 303 E. Chicago Avenue
> Chicago, IL 60611
> (312) 503-4140
>
> http://cam.facilities.northwestern.edu/
> http://nic.feinberg.northwestern.edu/
>


-- 



George McNamara, Ph.D.
Houston, TX 77054
https://www.linkedin.com/in/georgemcnamara