We provide here a general introduction on chlorophyll (Chl) a fluorescence, then we present our measurements on fast (< 1 s) induction curves (the so-called OJIP transients) on dark-adapted intact leaves of Arabidopsis thaliana, under five different light intensities [in the range of ~ 500 to ~ 3,000 µmol(photons) m‒2 s‒1] using two different instruments: Handy PEA (Hansatech Instruments, UK; excitation light, 650 nm) and FluorPen (model FP-110; Photon Systems Instruments, The Czech Republic; excitation light, 470 nm). We then discuss the observed differences in the OJIP curves, as well as in Fo (F20μs, F50μs, or the extrapolated Ft→0), FP (the peak), and the ratios FP/Fo, and Fv (= FP ‒ Fo)/FP in terms of differences in excitation light intensity and absorptance (or absorbance) of the excitation light by the leaves, and other factors, as well as the data available in the literature. We suggest that such measurements be accompanied, in the future, by parallel measurements on Chl a fluorescence imaging, an area pioneered by Hartmut K. Lichtenthaler., B. Padhi, G. Chauhan, D. Kandoi, A. Stirbet, B. C. Tripathy, G. Govindjee., and Obsahuje bibliografické odkazy
Transients of chlorophyll fluorescence in photosynthetic objects are often measured using short pulses of exciting radiation, which has recently been employed to capture kinetic images of fluorescence at the macroscopic level. Here we describe an instrument introducing this principle to recording of two dimensional fluorescence transients in microscopic objects. A modified fluorescence microscope is equipped with a CCD camera intensified by a micro-channel plate image amplifier. The microscopic field is irradiated simultaneously by three types of radiation: actinic radiation, saturating flashes, and pulsed measuring radiation. The measuring pulses are generated by a light-emitting diode and their duration is between 10 to 250 µs. The detection of fluorescence images (300×400 pixels, 8 bit) has a maximum time resolution of 40 ms and is gated in synchrony with the exciting pulses. This allows measuring on a background of a continuous actinic radiation up to irradiance that can elicit the maximal fluorescence yield (FM). On the other hand, the integral irradiance of the objects by the measuring radiation is very low, e.g., 0.08 µmol m-2 s-1 at 0.05 µm spatial resolution and 0.006 µmol m-2 s-1 at 4 µm spatial resolution. This allows a reliable recording of F0 even in very short time intervals (e.g., 5×80 ms). The software yields fluorescence kinetic curves for objects in user-selected areas as well as complete false-colour maps of the essential fluorescence kinetics parameters (FM, FO, FV, FV/FM, etc.) showing a two-dimensional distribution of their values. Several examples demonstrate that records of fluorescence kinetics can be obtained with a reasonable signal-to-noise ratio with all standard microscope objectives and with object sizes reaching from segments of leaf tissue to individual algal cells or chloroplasts. and H. Küpper ... [et al.].
The recognition of aquatic organisms plays a crucial role in the monitoring of the pollution and for the adoption of rapid preventive actions. A compact microscopic optical imaging system is proposed in order to acquire and treat the multibands fluorescence of several pigments in phytoplankton organisms. Two algorithms for automatic recognition of phytoplankton were proposed with a minimum number of calibration parameters. The first algorithm provides a morphological recognition based on "watershed" segmentation and Fourier descriptors, while the second one builds fluorescence pigment images by "k-means" partition of intensity ratios. The operation of these algorithms was illustrated by the study of two different organisms: a cyanobacteria (Dolichospermum sp.) and an alga (Cladophora sp.). The family and the genus of these organisms were then classified into a database which is independent of the size, the orientation and the position of the specimens in the images., M. Lauffer, F. Genty, S. Margueron, J. L. Collette., and Obsahuje bibliografii
The effect of high irradiance (HI) during desiccation and subsequent rehydration of the homoiochlorophyllous desiccation-tolerant shade plant Haberlea rhodopensis was investigated. Plants were irradiated with a high quantum fluence rate (HI; 350 µmol m-2 s-1 compared to ca. 30 µmol m-2 s-1 at the natural rock habitat below trees) and subjected either to fast desiccation (tufts dehydrated with naturally occurring thin soil layers) or slow desiccation (tufts planted in pots in peat-soil dehydrated by withholding irrigation). Leaf water content was 5 % of the control after 4 d of fast and 19 d of slow desiccation. Haberlea was very sensitive to HI under all conditions. After 19 d at HI, even in well-watered plants there was a strong reduction of rates of net photosynthesis and transpiration, contents of chlorophyll (Chl) and carotenoids, as well as photosystem 2 activity (detected by the Chl fluorescence ratio RFd). Simultaneously, the blue/red and green/red fluorescence ratios increased considerably suggesting increased synthesis of polyphenolic compounds. Desiccation of plants in HI induced irreversible changes in the photosynthetic apparatus and leaves did not recover after rehydration regardless of fast or slow desiccation. Only young leaves survived desiccation. and K. Georgieva, S. Lenk, C. Buschmann.