Detecting Signals at the Single Molecule Level: Pioneering
Achievements in Microscopy
Recent advances have led to such remarkable improvements in
fluorescence lifetime imaging microscopy's (FLIM) capacity for
contrast and sensitivity that researchers can now employ it to
detect signals at the single molecule level. FLIM also offers the
additional benefit of independence from fluorophore concentration
and excitation intensity. Moreover, its unique sensitivity makes it
an excellent reporter of conformational changes and of variations
in the molecular surroundings of biological molecules.
Most of this improvement and discovery have occurred during the
past decade, and, to date, information that would benefit a broad
range of researchers remains scattered in the literature. Edited by
two of the top pioneers in the field, FLIM Microscopy in Biology
and Medicine presents the fundamentals of FLIM along with a number
of advanced considerations so that a wider audience can appreciate
recent and potential improvements that make it such a valuable
tool.
New Opportunities for Biomedical Researchers... New Challenges
for Microscopy Researchers
Discussion sections in all the chapters clearly show the
challenges for implementing FLIM for various applications. Certain
chapters discuss limits on the number of photons required for
highly accurate lifetime determinations, as well as the accuracy
with which multiple, closely associated lifetime components can
reliably be determined. Such considerations are important for the
user when he or she is selecting the most advantageous method of
FLIM to use for a particular application.
While this book provides an introduction for those new to FLIM,
it gathers a wealth of material to enhance the work of experts
involved in pioneering technological improvements, as well as those
research opportunities in this unique and promising area of
microscopy.
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