scholarly journals Dye distance mapping using waveguide evanescent field fluorescence microscopy and its application to cell biology

2014 ◽  
Vol 8 (10) ◽  
pp. 826-837 ◽  
Author(s):  
Frederik Fleissner ◽  
Michael Morawitz ◽  
S. Jeffrey Dixon ◽  
Uwe Langbein ◽  
Silvia Mittler
Keyword(s):  
Fluorescence Microscopy ◽  
Cell Biology ◽  
Evanescent Field ◽  
Distance Mapping

Waveguide evanescent field fluorescence microscopy: Thin film fluorescence intensities and its application in cell biology

2008 ◽  
Vol 92 (23) ◽  
pp. 233503 ◽  
Author(s):  
Abdollah Hassanzadeh ◽  
Michael Nitsche ◽  
Silvia Mittler ◽  
Souzan Armstrong ◽  
Jeff Dixon ◽  
...  
Keyword(s):  
Thin Film ◽  
Fluorescence Microscopy ◽  
Cell Biology ◽  
Evanescent Field

Application of FRAP and digitized fluorescence microscopy to problems in cell membrane dynamics

1988 ◽  
Vol 46 ◽  
pp. 118-119
Author(s):  
K. Jacobson ◽  
A. Ishihara ◽  
B. Holifield ◽  
F. Zhang
Keyword(s):  
Fluorescence Microscopy ◽  
Cell Biology ◽  
Extended Period ◽  
Dynamic Structure ◽  
Living Cells ◽  
Membrane Dynamics ◽  
Molecular Cell Biology ◽  
Image Averaging ◽  
Single Living Cells ◽  
Single Living

Our laboratory is concerned with understanding the dynamic structure of the plasma membrane with particular reference to the movement of membrane constituents during cell locomotion. In addition to the standard tools of molecular cell biology, we employ both fluorescence recovery after photo- bleaching (FRAP) and digitized fluorescence microscopy (DFM) to investigate individual cells. FRAP allows the measurement of translational mobility of membrane and cytoplasmic molecules in small regions of single, living cells. DFM is really a new form of light microscopy in that the distribution of individual classes of ions, molecules, and macromolecules can be followed in single, living cells. By employing fluorescent antibodies to defined antigens or fluorescent analogs of cellular constituents as well as ultrasensitive, electronic image detectors and video image averaging to improve signal to noise, fluorescent images of living cells can be acquired over an extended period without significant fading and loss of cell viability.

scholarly journals Differential equation methods for simulation of GFP kinetics in non–steady state experiments

2018 ◽  
Vol 29 (6) ◽  
pp. 763-771 ◽  
Author(s):  
Robert D. Phair
Keyword(s):  
Differential Equation ◽  
Steady State ◽  
Fluorescence Microscopy ◽  
Cell Biology ◽  
Fluorescent Proteins ◽  
Quantitative Information ◽  
Tracer Kinetics ◽  
Mathematical Framework ◽  
Experimental Protocols ◽  
Tracer Kinetic

Genetically encoded fluorescent proteins, combined with fluorescence microscopy, are widely used in cell biology to collect kinetic data on intracellular trafficking. Methods for extraction of quantitative information from these data are based on the mathematics of diffusion and tracer kinetics. Current methods, although useful and powerful, depend on the assumption that the cellular system being studied is in a steady state, that is, the assumption that all the molecular concentrations and fluxes are constant for the duration of the experiment. Here, we derive new tracer kinetic analytical methods for non–steady state biological systems by constructing mechanistic nonlinear differential equation models of the underlying cell biological processes and linking them to a separate set of differential equations governing the kinetics of the fluorescent tracer. Linking the two sets of equations is based on a new application of the fundamental tracer principle of indistinguishability and, unlike current methods, supports correct dependence of tracer kinetics on cellular dynamics. This approach thus provides a general mathematical framework for applications of GFP fluorescence microscopy (including photobleaching [FRAP, FLIP] and photoactivation to frequently encountered experimental protocols involving physiological or pharmacological perturbations (e.g., growth factors, neurotransmitters, acute knockouts, inhibitors, hormones, cytokines, and metabolites) that initiate mechanistically informative intracellular transients. When a new steady state is achieved, these methods automatically reduce to classical steady state tracer kinetic analysis.

Microscopy and Cell Biology: New Methods and New Questions

2019 ◽  
Vol 70 (1) ◽  
pp. 199-218
Author(s):  
Joshua D. Morris ◽  
Christine K. Payne
Keyword(s):  
Biological Sciences ◽  
Fluorescence Microscopy ◽  
Spatial Resolution ◽  
Cell Biology ◽  
Molecular Level ◽  
Label Free ◽  
Imaging Methods ◽  
New Methods ◽  
Multiple Imaging ◽  
Health And Disease

Understanding the cellular basis of human health and disease requires the spatial resolution of microscopy and the molecular-level details provided by spectroscopy. This review highlights imaging methods at the intersection of microscopy and spectroscopy with applications in cell biology. Imaging methods are divided into three broad categories: fluorescence microscopy, label-free approaches, and imaging tools that can be applied to multiple imaging modalities. Just as these imaging methods allow researchers to address new biological questions, progress in biological sciences will drive the development of new imaging methods. We highlight four topics in cell biology that illustrate the need for new imaging tools: nanoparticle-cell interactions, intracellular redox chemistry, neuroscience, and the increasing use of spheroids and organoids. Overall, our goal is to provide a brief overview of individual imaging methods and highlight recent advances in the use of microscopy for cell biology.

scholarly journals Correlating Fluorescence Microscopy with Electron Microscopy

2004 ◽  
Vol 12 (1) ◽  
pp. 3-7
Author(s):  
Stephen W. Carmichael ◽  
Jon Charlesworth
Keyword(s):  
Electron Microscopy ◽  
Transmission Electron Microscopy ◽  
Fluorescence Microscopy ◽  
Fluorescent Probes ◽  
Cell Biology ◽  
Microscopic Image ◽  
Electron Microscopic ◽  
Transmission Electron ◽  
Electron Microscopic Image ◽  
Auto Fluorescence

The use of fluorescent probes is becoming more and more common in cell biology. It would be useful if we were able to correlate a fluorescent structure with an electron microscopic image. The ability to definitively identify a fluorescent organelle would be very valuable. Recently, Ying Ren, Michael Kruhlak, and David Bazett-Jones devised a clever technique to correlate a structure visualized in the light microscope, even a fluorescing cell, with transmission electron microscopy (TEM).Two keys to the technique of Ren et al are the use of grids (as used in the TEM) with widely spaced grid bars and the use of Quetol as the embedding resin. The grids allow for cells to be identified between the grid bars, and in turn the bars are used to keep the cell of interest in register throughout the processing for TEM. Quetol resin was used for embedding because of its low auto fluorescence and sectioning properties. The resin also becomes soft and can be cut and easily peeled from glass coverslips when heated to 70°C.

Rigorous electromagnetic theory for waveguide evanescent field fluorescence microscopy

2018 ◽  
Vol 57 (30) ◽  
pp. 9129
Author(s):  
Abdollah Hassanzadeh ◽  
Shabbo Saedi ◽  
Mohammadbagher Mohammadnezhad ◽  
Salah Raza Saeed
Keyword(s):  
Fluorescence Microscopy ◽  
Electromagnetic Theory ◽  
Evanescent Field

scholarly journals Accuracy and precision in quantitative fluorescence microscopy

2009 ◽  
Vol 185 (7) ◽  
pp. 1135-1148 ◽  
Author(s):  
Jennifer C. Waters
Keyword(s):  
Fluorescence Microscopy ◽  
Cell Biology ◽  
Imaging System ◽  
Resonance Energy Transfer ◽  
Resonance Energy ◽  
Digital Cameras ◽  
Accuracy And Precision ◽  
Fluorescent Molecules ◽  
Quantitative Fluorescence ◽  
Biology Research

The light microscope has long been used to document the localization of fluorescent molecules in cell biology research. With advances in digital cameras and the discovery and development of genetically encoded fluorophores, there has been a huge increase in the use of fluorescence microscopy to quantify spatial and temporal measurements of fluorescent molecules in biological specimens. Whether simply comparing the relative intensities of two fluorescent specimens, or using advanced techniques like Förster resonance energy transfer (FRET) or fluorescence recovery after photobleaching (FRAP), quantitation of fluorescence requires a thorough understanding of the limitations of and proper use of the different components of the imaging system. Here, I focus on the parameters of digital image acquisition that affect the accuracy and precision of quantitative fluorescence microscopy measurements.

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