Label‐free bioanalysis of Leishmania infantum using refractive index tomography with partially coherent illumination

In this work, we report the use of refractive index (RI) tomography for quantitative analysis of unstained DH82 cell line infected with Leishmania infantum. The cell RI is reconstructed by using a modality of optical diffraction tomography technique that employs partially coherent illumination, thus enabling inherent compatibility with conventional wide‐field microscopes. The experimental results demonstrate that the cell dry mass concentration (DMC) obtained from the RI allows for reliable detection and quantitative characterization of the infection and its temporal evolution. The RI provides important insight for studying morphological changes, particularly membrane blebbing linked to an apoptosis (cell death) process induced by the disease. Moreover, the results evidence that infected DH82 cells exhibit a higher DMC than healthy samples. These findings open up promising perspectives for clinical diagnosis of Leishmania.     

Nanoscale Subsynaptic Domains Underlie the Organization of the Inhibitory Synapse

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Floral development in Scutellaria pinnatifida (Lamiaceae): the ontogenetic basis for sepal reduction

The Scutellaria is a Labiatae genus (subfamily Scutellarioideae) with a highly specialised floral structure. The genus is characterised by a peculiar two‐lobed calyx with a projecting appendage, named the scutellum. Here, we present a detailed analysis of floral development, using epi‐illumination light microscopy, to clarify open questions about its floral organisation. Floral whorls appeared in an acropetal sequence, with a marked temporal overlap of petal and stamen appearance. Organ appearance in each whorl proceeded unidirectionally from the abaxial to the adaxial side. Significant developmental features included the formation of the scutellum, reduction of sepal lobes and formation of a three‐lobed nectary disc. Our study revealed that both loss of organ initiation and fusion of primordia are responsible for the reduction in sepal members in Scutellaria. The nectary structure was markedly different from most other studied Lamiaceae.     

雪灾干扰后林冠开阔度对黄心树幼苗更新的影响

幼苗更新是森林更新的关键过程,而林窗对于植物幼苗更新、生长及最终存活具有重要影响。极端气候事件会使森林在短时间内形成大量林窗,从而显著影响林内幼苗更新和存活。本研究以2015年雪灾后云南省哀牢山中山湿性常绿阔叶林优势种黄心树(Machilus bombycina)幼苗为对象,对194个1 m×1 m样方中的幼苗生长和死亡动态进行为期4年的监测,利用线性混合模型(LMM)和广义线性混合模型(GLMM),分析林冠开阔度对幼苗高度相对生长率、叶片相对增长率、新增率和死亡率的影响,研究了雪灾干扰后的林冠开阔度对植物幼苗更新的影响。结果表明:(1)林冠开阔度随时间推移逐渐变小(20.07%~9.97%),且速度由快到慢;(2)2015-2016年,已有幼苗高度相对生长率与林冠开阔度呈显著正相关(P<0.05),其余年份两者间无显著相关性;2016-2017年已有幼苗叶片相对增长率与林冠开阔度呈显著正相关(P<0.05),其余年份两者间无显著相关性;(3)林冠开阔度与幼苗新增率和死亡率均无显著相关性。本研究表明,雪灾后造成的林冠开阔度增加有利于黄心树幼苗生长。     

白云山国家森林公园不同人为干扰强度的群落冠层结构和光照特征

为探究森林冠层结构与林下光照的变化规律及其相关性,在河南省白云山国家森林公园选取人工林、择伐林、皆伐林和老龄林建立四块面积为1 hm²（100 m×100 m）的固定监测样地,利用半球面影像技术获取冠层结构及林下光照数据。研究发现：1）随着人为干扰强度降低,冠层覆盖度与叶面积指数呈增加趋势,林下散射辐射与直接辐射呈减少趋势;2）择伐林冠层覆盖度与叶面积指数最大,平均叶倾角与透光比最小,林下光照（直接辐射、散射辐射）与冠层覆盖度和叶面积指数都呈显著负相关,林下散射辐射与冠层覆盖度和叶面积指数的负相关关系最强;3）处于不同干扰强度的群落由于冠层结构的差异,形成了不同群落内特定的光照环境。研究结果丰富了暖温带-亚热带生态过渡区森林冠层结构与林下光照动态变化研究资料,同时也为该区域森林的恢复与重建等提供科学的依据。     

Detection of single quantum dots in model organisms with sheet illumination microscopy

Single-molecule detection and tracking is important for observing biomolecule interactions in the microenvironment. Here we report selective plane illumination microscopy (SPIM) with single-molecule detection in living organisms, which enables fast imaging and single-molecule tracking and optical penetration beyond 300 μm. We detected single nanocrystals in Drosophila larvae and zebrafish embryo. We also report our first tracking of single quantum dots during zebrafish development, which displays a transition from flow to confined motion prior to the blastula stage. The new SPIM setup represents a new technique, which enables fast single-molecule imaging and tracking in living systems.     

Drosophila pupal macrophages – A versatile tool for combined ex vivo and in vivo imaging of actin dynamics at high resolution

Molecular understanding of actin dynamics requires a genetically traceable model system that allows live cell imaging together with high-resolution microscopy techniques. Here, we used Drosophila pupal macrophages that combine many advantages of cultured cells with a genetic in vivo model system. Using structured illumination microscopy together with advanced spinning disk confocal microscopy we show that these cells provide a powerful system for single gene analysis. It allows forward genetic screens to characterize the regulatory network controlling cell shape and directed cell migration in a physiological context. We knocked down components regulating lamellipodia formation, including WAVE, single subunits of Arp2/3 complex and CPA, one of the two capping protein subunits and demonstrate the advantages of this model system by imaging mutant macrophages ex vivo as well as in vivo upon laser-induced wounding.     

补光对越冬灰仓鼠体重、主要器官指数及精子数量的影响

目的研究补光对灰仓鼠(Cricetulus migratorus)越冬时脏器器官的影响。方法室温下将其分为自然光照组和补充光照组,分别在第0、30、48和150天时测定动物的体重和脏器器官的重量,应用涂片法检查精子的数量。结果自然光照时,精子数量在冬至时变化不大,但冬至后第18天,灰仓鼠的睾丸系数降到最低值,精子数量仅为422枚,第150天时恢复到正常繁殖状态,精子数量达到7440枚。其他脏器指数在冬至时最高,第150天时最低。补充光照后,雌雄鼠体重高峰值时间均比自然光照组提前100 d,各器官指数变化与体重变化情况相类似,但睾丸和子宫系数维持稳定,精子数量未发现减少。结论在自然光照时越冬灰仓鼠的脏器指数逐渐下降,第150天时脏器指数达至最高值,睾丸指数在冬至后第18天下降到最低值,处于非繁殖状态。补充光照后,越冬灰仓鼠的精子数量和子宫指数变化不大,一直维持繁殖状态。     

Fluorescence Microscopy Gets Faster and Clearer: Roles of Photochemistry and Selective Illumination

Significant advances in fluorescence microscopy tend be a balance between two competing qualities wherein improvements in resolution and low light detection are typically accompanied by losses in acquisition rate and signal-to-noise, respectively. These trade-offs are becoming less of a barrier to biomedical research as recent advances in optoelectronic microscopy and developments in fluorophore chemistry have enabled scientists to see beyond the diffraction barrier, image deeper into live specimens, and acquire images at unprecedented speed. Selective plane illumination microscopy has provided significant gains in the spatial and temporal acquisition of fluorescence specimens several mm in thickness. With commercial systems now available, this method promises to expand on recent advances in 2-photon deep-tissue imaging with improved speed and reduced photobleaching compared to laser scanning confocal microscopy. Superresolution microscopes are also available in several modalities and can be coupled with selective plane illumination techniques. The combination of methods to increase resolution, acquisition speed, and depth of collection are now being married to common microscope systems, enabling scientists to make significant advances in live cell and in situ imaging in real time. We show that light sheet microscopy provides significant advantages for imaging live zebrafish embryos compared to laser scanning confocal microscopy.     

Effect of Temperature and Illumination on Growth and Reproduction of the Green Alga Ulva fenestrata

The combined effect of temperature (5, 10, 15, and 20°C) and illumination (40 and 60 mE/(m² s)) on growth and reproduction of the green marine alga Ulva fenestrata P. et R. from the sublittoral zone of Amursky Bay, Sea of Japan, was studied in the laboratory environment in the months April–July, 2000. It was demonstrated that the temperature of 5°C and illumination of 40 mE/(m² s) are the most favorable for maintaining the vegetative mass of the algae. A water temperature of 10°C and illumination of 40 mE/(m² s) are the optimum conditions for vegetative growth of U. fenestrata thalli. A temperature decrease and increase by 5°C reduces the growth rate on average by 30%. Sporo- and gametogenesis in U. fenestrata are the most regular (every 10 days) and occupy the greatest disk area at a water temperature of 15°C and illumination of 40 mE/(m² s). Vegetative growth of thalli is sharply inhibited at the stage of cell preparation to gametogenesis a day before the beginning of gamete formation.