PubMed · 14817885
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A LeSAGE. 1951. [The cell; its intelligence].. https://pubmed.ncbi.nlm.nih.gov/14817885/
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BACKGROUND AND OBJECTIVES: Laser-activated micro- and nano-bubbles (LAB) in cells may be used as universal and sensitive non-toxic probes for measuring functional properties of individual cells. STUDY DESIGN/MATERIALS AND METHODS: Such bubbles can be detected and imaged by microscopy and flow cytometry. LABs in living blood and tumor cells were induced by pulsed (532 nm, 10 nanoseconds) laser radiation and detected by the thermal lens optical method. RESULTS: Registered lifetime and maximal diameter of the studied LABs varied within the ranges of 0.02-10 microseconds and 0.44-100 microm, respectively. LAB parameters, thresholds and probabilities, were found to depend upon the physiological state of cells. Specificity and sensitivity of LAB cytometry were increased due to the use of light-absorbing nanoparticles conjugated to specific monoclonal antibodies. CONCLUSIONS: LAB were found to be the universal phenomena that can be used for sensitive and non-invasive monitoring of any individual cell, intact or nanoparticle-treated.
Focus maintenance is a challenging problem in multidimensional wide-field microscopy. Most automated microscopes use software algorithms, which are applied to z-sections of the object, to select for the plane with the best signal to noise ratio. When applied automatically in multidimensional imaging applications, auto focus routines significantly increase light exposure and can become cytotoxic if applied too frequently. In addition, automated focusing procedures can readily focus on unwanted high contrast objects. By labelling a defined position with a fluorescent marker, we were able to separate the focusing procedure from the actual image acquisition positions and therefore overcome some of the major drawbacks of routine auto focus procedures. To implement this method in a multidimensional acquisition experiment, we created a visual basic-based program, which is run prior to each image acquisition. This technique allows tight control of focus whilst keeping light toxicity in live cell imaging experiments to a minimum.
Adaptation and behavior are characteristics of life which are fundamentally dynamic. If we want to model the living cell we have to describe it as a dynamic system. Typical dynamic models are based on quantitative differential equations requiring very detailed kinetic knowledge. Alternative modeling techniques for less fine-grained information are better suited to available functional genomics data. As such, constraint-based techniques and qualitative modeling have proven themselves to be valid approaches in cell biology. These approaches offer formal support to check the consistency of molecular networks against phenotypic observations in the light of dynamic systems.