The scale bar dialog will open and a scale bar will appear on your image. In the Analyze/Tools menu select Scale Bar. From basic documentation to advanced life science research LAS X directly navigates you to brilliant imaging5) Now open the image you want to add a scale bar to. Leica Application Suite X (LAS X) is the one software platform for all Leica microscopes: It integrates confocal, widefield, stereo, super-resolution, and light-sheet instruments from Leica Microsystems.
![]() 2014 Sofroniew 2015 Burda et al. 2009 Jinno 2011 Nash et al. The idea that astrocytic process arbors vary consistently among species, brain regions, and physiological states is widely accepted (Bushong et al. 2012 Khakh and Sofroniew 2015). 2015 Yang and Wang 2015 Burda et al. 2012 Hol and Pekny 2015 Sofroniew 2015 Kulkarni et al. For instance, in some pathologies astrocytes undergo a dramatic increase in the process thickness and ramification named astrogliosis (Nash et al. Moreover, astrocyte morphology is largely affected under pathological conditions. The choice between options is normally based upon both the level of detail needed to answer the scientific question and the resources that are available. Currently, there are multiple tools available to analyze and quantify the extremely complex astrocyte morphology (Parekh and Ascoli 2013 Kulkarni et al. Get data-driven recommendations and read in-depth descriptions to find the right toolsAssessing astrocyte structure and morphology requires complementary approaches to assess varying levels of detail (main process arbor versus fine process). 2014 Khakh and Sofroniew 2015 Heller and Rusakov 2015).Discover the best business apps in 2021 and check over 1 million verified ratings & reviews. Such structural alterations in astrocytes were shown to have functional consequences (Haber et al. 2012 Hol and Pekny 2015 Sofroniew 2015 Kulkarni et al. The brain samples were stained against glial fibrillary acidic protein (GFAP), which is the intermediate filament protein that is used extensively as a specific marker for astrocyte main processes, whose expression is tightly related to morphological alterations (Wilhelmsson et al. This method allowed to semi-automatically screen the astrocyte structure in a large number of samples, in a simple, effective and costless manner. 2011), to reconstruct tridimensional arbors of astrocytic main processes. Taking these factors into account, we applied the open-source tool, Simple Neurite Tracer (SNT) (Longair et al. On the other hand, commercially available tools are rather user-friendly, yet they are costly and time-consuming. ![]() The acquisition settings were the following: scanning speed, 4 µs/px pinhole aperture, 110 µm GFAP, excitation = 559 nm, emission = 618 nm DAPI, excitation = 405 nm, emission = 461 nm pinhole aperture = 110 µm. 3a), to work with a similar image resolution. 0.90 dry field size 317.13 × 317.13 µm 0.31 µm/px Fig. 2, 3, 4, 5) and 1024 × 1024 px using a 40× objective (UPlanSApo, N.A. 1.42 oil field size 211.51 × 211.51 µm 0.33 µm/px Figs. A resolution of 640 × 640 px was achieved using a 60× objective (PlanApo N, N.A. C At process complexity level, no differences were found between magnifications. B Similar results of astrocytic morphology were obtained from reconstruction at 40× and 60× magnification, both for total length and number of processes. A Representative GFAP–DAPI staining micrograph (max projection) under 40× and 60× magnification scale bar 50 µm. What is the microsoft office for macB Representative images of 3D astrocytic morphology reconstruction obtained from different users with SNT. A Representative GFAP–DAPI staining micrograph (max projection) under 60× magnification, showing detailed astrocytic morphology scale bar 50 µm. * p < 0.05 ** p < 0.01 different from 1 µm # p < 0.05 and # p < 0.01 different from 2 µmSimple Neurite Tracer is a reliable tool to reconstruct astrocyte process arbors. Data plotted as mean ± SEM. F Sholl analysis confirms loss of detailed information regarding process arbor complexity at higher Z-step interslice intervals. E No statistically significant differences were found between 1 and 2 µm Z-step interval 3 and 4 µm leads to a significant loss of detail in astrocytic morphology and unreliable measures of total length and number of processes. Sem Software Image Tif Scale Bar Full Size ImageThe selection criteria used (examples in Fig. G Sholl analysis confirms similarity between reconstructed astrocytic morphologies at a detailed level of arbor complexityFull size image Astrocyte selection criteria for reconstructionAstrocytes were readily identified by their characteristic GFAP-positive bushy shape, displaying thicker processes around the DAPI-stained nucleus. F Astrocytic reconstructions obtained from AutoNeuron and SNT confirm similarity between total length and number of processes. E Representative GFAP + cell micrograph (max projection) and its respective reconstruction obtained from AutoNeuron and SNT scale bar 50 µm. D Sholl analysis shows similarity of process arbor complexity of reconstructed astrocytes from different SNT users.
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