Warm Cells, Hot Mitochondria: Achievements and Problems of Ultralocal Thermometry.

Temperature is a crucial regulator of the rate and direction of biochemical reactions and cell processes. The recent data indicating the presence of local thermal gradients associated with the sites of high-rate thermogenesis, on the one hand, demonstrate the possibility for the existence of "thermal signaling" in a cell and, on the other, are criticized on the basis of thermodynamic calculations and models. Here, we review the main thermometric techniques and sensors developed for the determination of temperature inside living cells and diverse intracellular compartments. A comparative analysis is conducted of the results obtained using these methods for the cytosol, nucleus, endo-/sarcoplasmic reticulum, and mitochondria, as well as their biological consistency. Special attention is given to the limitations, possible sources of errors and ambiguities of the sensor's responses. The issue of biological temperature limits in cells and organelles is considered. It is concluded that the elaboration of experimental protocols for ultralocal temperature measurements that take into account both the characteristics of biological systems, as well as the properties and limitations of each type of sensor is of critical importance for the generation of reliable results and further progress in this field.

Comparison of chlorin-e6 detection efficiency by video systems with excitation wavelengths of 405nm and 635nm.

BACKGROUND: Fluorescence diagnostics with two different wide field-of-view imaging systems with fluorescence excitation at 405 nm and 635 nm, respectively, were compared. Both systems include fluorescence quantification and experimental geometry normalization algorithms. METHODS: A newly developed system with an excitation wavelength of 405 nm was tested on intralipid fluorescent tumor phantoms with chlorin-e6. Both, this new system and a second existing system with an excitation wavelength of 635 nm, were used for fluorescent diagnosis in six patients with basal cell carcinoma and cancer of the oral mucosa. For PDT, a red diode laser with a wavelength of 660 nm was used for all 6 patients. One patient received an additional irradiation using the red LED source of the new system RESULTS: The boundaries of the lesions and the fluorescence intensity were successfully determined by both video systems. CONCLUSIONS: Both fluorescence imaging approaches showed comparable contrast between diseased and healthy tissues. For oral mucosal cancer, a system with violet fluorescence excitation, bispectral frame analysis, and time-resolved background suppression showed better contrast between the tumor and normal tissue and effective elimination of autofluorescence. Moreover, both systems provided efficient quantification of fluorescence and gave fluorescence indices that were weakly dependent on the distance between the device and the tissue.