Involvement of vitamin A in the photoperiodic induction of diapause in the spider mite Tetranychus urticae is demonstrated by rearing an albino mutant on a semi-synthetic diet with and without p-carotene or vitamin A

Abstract. Data are presented on the development from egg to adult of a wild-type and an albino strain of the spider mite Tetranychus urticae on a semi-synthetic liquid diet.The diet is a modification of the diet developed by Van der Geest et al. (1983).Apart from the standard diet without added carotenoids, diets supplemented with either β-carotene or vitamin A were tested.A high incidence of diapause was found in the wild-type strain under short-day conditions on all three diets.No diapause was found in the albino mutant under short days on the standard diet.Partial restoration of the photoperiodic response was obtained after addition of β-carotene to the diet, and full restoration was found after the addition of vitamin A.Diapause could be terminated in diet-reared diapausing females of both strains after a cold rest of 3 weeks at 4°C.The results lead to the conclusion that vitamin A or a derivative of vitamin A probably functions as the photoreceptor pigment for the photoperiodic induction of diapause in the spider mite.     

Toward absolute viability measurements for bacteria

We aim to develop a quantitative viability method that distinguishes individual quiescent from dead cells and is measured in time (ns) as a referenceable, comparable quantity. We demonstrate that fluorescence lifetime imaging of an anionic, fluorescent membrane voltage probe fulfills these requirements for Streptococcus mutans. A random forest machine-learning model assesses whether individual S. mutans can be correctly classified into their original populations: stationary phase (quiescent), heat killed and inactivated via chemical fixation. We compare the results to intensity using three models: lifetime variables (τ₁, τ₂ and p₁), phasor variables (G, S) or all five variables, with the five variable models having the most accurate classification. This initial work affirms the potential for using fluorescence lifetime of a membrane voltage probe as a viability marker for quiescent bacteria, and future efforts on other bacterial species and fluorophores will help refine this approach.     

Lightsheet fluorescence lifetime imaging microscopy with wide‐field time‐correlated single photon counting

We report on wide‐field time‐correlated single photon counting (TCSPC)‐based fluorescence lifetime imaging microscopy (FLIM) with lightsheet illumination. A pulsed diode laser is used for excitation, and a crossed delay line anode image intensifier, effectively a single‐photon sensitive camera, is used to record the position and arrival time of the photons with picosecond time resolution, combining low illumination intensity of microwatts with wide‐field data collection. We pair this detector with the lightsheet illumination technique, and apply it to 3D FLIM imaging of dye gradients in human cancer cell spheroids, and C. elegans.     

Lifetime fitness components in female colour morphs of a damselfly: density‐ or frequency‐dependent selection?

In many damselfly species mature females exhibit colour polymorphism: one female morph resembles the conspecific male (androchrome) while the others do not (gynochromes). Hypotheses for the maintenance of such polymorphisms differ mainly as to whether they are based on density- and/or frequency-dependent selection and on the nature of the frequency dependence. We collected lifetime fitness data (individual lifespan, number of copulations and number of ovipositions) for female morphs of the damselfly Ischnura elegans from 15 insectaries differing in population parameters (density, sex ratio and ratio of andro- to gynochromes). Both density and frequency affected a specific set of the studied fitness components. While morph frequency influenced lifespan, sex ratio influenced the number of copulations, and density affected lifespan and the number of ovipositions. Clearly, discrepancies among studies may be generated if the studied fitness components differ. Our final fitness estimate, the number of ovipositions, was only influenced by density, thereby not supporting frequency-based hypotheses. Contrary to expectation under the current density-based hypothesis, androchromes compared to gynochromes had a lower number of ovipositions at high density. We discuss our findings in the light of mechanisms maintaining the female polymorphism.  © 2005 The Linnean Society of London, Biological Journal of the Linnean Society , 2005, 86 , 515–523.     

Insect oviposition plasticity in response to host availability: the case of the tephritid fruit fly Bactrocera dorsalis

1. Insect oviposition behaviour is ecologically and physiologically plastic. For tephritid fruit flies, Bactrocera dorsalis Hendel, host availability varies spatially and temporally. Females are expected to adopt adaptive oviposition strategies to maximise lifetime reproductive fitness, including survival. Bactrocera dorsalis oviposition tactics in response to different host availabilities were investigated. 2. This study includes three treatments: (i) variable host densities (host density varied according to a fixed cycle from day to day over values of 1, 5, 10 and 20 hosts per cage), (ii) a fixed high host density (20 hosts per cage), and (iii) a fixed low host density (1 host per cage). 3. Daily egg‐laying number per female over the course of 27 days was entirely independent of host density and highly dependent on female age. As host availability increased, females accepted significantly more hosts, generally laid small egg clutches, and more broadly distributed the eggs. 4. Tephritid fruit flies adaptively adjusted egg clutches in ways that reflected the variability of host availability. Egg‐ and time‐limitation constraints appeared to drive these adjustments. Female egg maturation was triggered by oviposition activity and reflected marked lifetime trade‐offs. Such strategies involved specific time schedules for egg laying. 5.This study defined the oviposition plasticity of the tephritid fruit fly. These results have general implications for the behavioural ecology of insect herbivores and parasitoids.     

Quenched coumarin derivatives as fluorescence lifetime phantoms for NADH and FAD

Two-photon fluorescence lifetime imaging is a versatile laboratory technique in the field of biophotonics and its importance is also growing in the field of in vivo diagnostics for medical purposes. After years of experience in dermatology, endoscopic implementations of the technique are now posing new technical challenges. To develop, test, and compare instrumental solutions for this purpose suitable reference samples have been devised and tested. These reference samples can serve as reliable NADH- and FAD-mimicking optical phantoms for 2-photon fluorescence lifetime imaging, as they can be prepared relatively easily with reproducible and stable characteristics for this quite relevant diagnostic technique. The reference samples (mixtures of coumarin 1 and coumarin 6 in ethanol with suitable amounts of 4-hydroxy-TEMPO) have been tuned to exhibit spectral and temporal fluorescence characteristics very similar to those of NADH and FAD, the two molecules most frequently utilized to characterize cell metabolism.     

Mesoscopic fluorescence lifetime imaging: Fundamental principles,clinical applications and future directions

Fluorescence lifetime imaging (FLIm) is an optical spectroscopic imaging technique capable of real-time assessments of tissue properties in clinical settings. Label-free FLIm is sensitive to changes in tissue structure and biochemistry resulting from pathological conditions, thus providing optical contrast to identify and monitor the progression of disease. Technical and methodological advances over the last two decades have enabled the development of FLIm instrumentation for real-time, in situ, mesoscopic imaging compatible with standard clinical workflows. Herein, we review the fundamental working principles of mesoscopic FLIm, discuss the technical characteristics of current clinical FLIm instrumentation, highlight the most commonly used analytical methods to interpret fluorescence lifetime data and discuss the recent applications of FLIm in surgical oncology and cardiovascular diagnostics. Finally, we conclude with an outlook on the future directions of clinical FLIm.     

Intraoperative detection of IDH-mutant glioma using fluorescence lifetime imaging

Identifying isocitrate dehydrogenase (IDH)-mutation and glioma subtype during surgery instead of days later can aid in modifying tumor resection strategies for better survival outcomes. We report intraoperative identification of IDH-mutant glioma (N = 12 patients) with a clinically compatible fluorescence lifetime imaging (FLIm) device (excitation: 355 nm; emission spectral bands: 390/40 nm, 470/28 nm, 542/50 nm). The fluorescence-derived parameters were analyzed to study the optical contrast between IDH-mutant tumors and surrounding brain tissue. IDH-mutant oligodendrogliomas exhibited shorter lifetimes (3.3 ± 0.1 ns) than IDH-mutant astrocytomas (4.1 ± 0.1 ns). Both IDH-mutant glioma subtypes had shorter lifetimes than white matter (4.6 ± 0.4 ns) but had comparable lifetimes to cortex. Lifetimes also increased with malignancy grade within IDH-mutant oligodendrogliomas (grade 2: 2.96 ± 0.08 ns, grade 3: 3.4 ± 0.3 ns) but not within IDH-mutant astrocytomas. The current results support the feasibility of FLIm as a surgical adjuvant for identifying IDH-mutant glioma tissue.     

Fluorescence lifetime and rotational correlation time of bovine serum albumin-sodium dodecyl sulfate complex labeled with 1-dimethylaminonaphthalene-5-sulfonyl chloride: Effect of disulfide bridges in the protein on these fluorescence parameters

A fluorescent dye, 1-dimethylaminonaphthalene-5-sulfonyl chloride, was used to label bovine serum albumin (BSA), intact and disulfide bridges-cleaved. The fluorescence lifetime and fluorescence anisotropy of the adducts in sodium dodecyl sulfate (SDS) solutions were studied by the nanosecond fluorescence depolarization method. The volume of equivalent sphere (V _e) was calculated to be 2.1×10^–19 cm³ for BSA with the intact disulfide bridges from the rotational correlation time. The value ofV _e was 4.4×10^–19 cm³ for the disulfide bridges-cleaved BSA. With an increase in SDS concentration, the rotational correlation time of the intact BSA became longer, while that of the disulfide bridges-cleaved BSA became shorter. This suggests that upon the binding of SDS, the total volume of the intact BSA increases while the expanded state of the protein, caused by the cleavage of the disulfide bridges, becomes compact.     

Metabolic transcriptomics dictate responses of cone photoreceptors to retinitis pigmentosa

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