Utilizing a custom-built, on-stage incubator-combined, two-photon excitation fluorescence (TPEF) and second
harmonic generation (SHG) imaging system, we observed new-sarcomere addition in rat neonatal cardiomyocytes
during 10 hours of on-stage incubation. This addition occurred at one end of an existing myofibril, the sides of
existing myofibrils, and at the interstice of several separated myofibrils; in the cases of the latter two, we observed
mature myofibrils acting as templates. We found that during sarcomeric addition, myosin filaments are assembled
onto the premyofibril laterally. This lateral addition, which proceeds stepwise along the axial direction, plays an
important role in the accumulation of Z-bodies to form mature Z-disks and in the regulation of sarcomeric length
during maturation.
We developed a hybrid two-photon excitation fluorescence-second harmonic generation (TPEF-SHG) imaging system with an on-stage incubator for long-term live-cell imaging. Using the imaging system, we observed the addition of new sarcomeres during myofibrillogenesis while a cardiomyocyte was spreading on the substrate. The results suggest that the TPEF-SHG imaging system with an on-stage incubator is an effective tool for investigation of dynamic myofibrillogenesis.
Laser guidance is the technique that uses a weakly convergent laser beam to trap particles radially in the center of the
beam and simultaneously propel them along the beam propagation axis with a travelling distance over millimeters. In
this paper, we describe the applications of laser guidance to detect different cell types, including those of phenotypically
transformed or gene-modified cells, especially for situations in which fluorescent markers used in flow cytometry for cell
detection are not available or their application is contraindicated by clinical restriction. The optical force, which
determines the guidance speed of the cell, is dependent on the characteristics, such as size, shape, composition and
refractive index, of the cell being guided. Therefore, by measuring the guidance speed of the cell along the laser beam,
cells with different properties can be effectively distinguished. We report two experimental results: 1) the laser-guidance
system could significantly distinguish the metastatic cancer cell type 4T1 from its non-metastatic counterpart 4T07,
which could not be achieved by using a high magnification microscope; 2) The laser-guidance experiment demonstrated
that only one gene modification between L-10 and TC-1 cells resulted in ~40% difference in guidance speed. These
experimental data indicate that laser guidance can be used to detect subtle differences between sub-cell types.
Access to the requested content is limited to institutions that have purchased or subscribe to SPIE eBooks.
You are receiving this notice because your organization may not have SPIE eBooks access.*
*Shibboleth/Open Athens users─please
sign in
to access your institution's subscriptions.
To obtain this item, you may purchase the complete book in print or electronic format on
SPIE.org.
INSTITUTIONAL Select your institution to access the SPIE Digital Library.
PERSONAL Sign in with your SPIE account to access your personal subscriptions or to use specific features such as save to my library, sign up for alerts, save searches, etc.