WT host mice were anesthetized by intraperitoneal injection of xylazine (12.5 mg/kg) and ketamine hydrochloride (125 mg/kg) (Sanofi, Mexico-City, Mexico). The inguinal lymph node was then inoculated with 100 μl CXCL13 (25 ng/ml) (PeproTech) to promote cell recruitment. One hour later, 1 × 107 Hoechst 33342-labeled, LPS + IL-4 activated B cells were injected via the cannulated carotid artery. Venules of the inguinal lymph node were recorded using an intravital upright microscope (Axioscope, Model A1, Zeiss, Jena, Germany) with a 40 × and 0.75 saline immersion objective (Zeiss). Videos and images were analyzed using ImageJ (NIH, Bethesda, MD. USA) and Zen Blue Edition 2.5 software (Zeiss, Microscopy). The venules' diameter, the number of adherent cells, the number of transmigrated cells, and the cells' velocity were measured using ImageJ. Cell flux, blood flow, and rolling velocity were analyzed by Zen Blue Edition 2.5 software (Zeiss, Microscopy). This methodology was carried out following a previously published protocol32 .
Zen 2 blue edition
Manufactured by Zeiss
Sourced in Germany, Japan
ZEN 2 blue edition is a microscopy software platform developed by Zeiss. It provides a comprehensive set of tools for image acquisition, processing, and analysis. The software is designed to work seamlessly with Zeiss microscopes and offer a user-friendly interface for researchers and technicians.
Automatically generated - may contain errors
Lab products found in correlation
Axio observer z1, by Zeiss (9 mentions)
Axiocam hr, by Zeiss (3 mentions)
M0851, by Agilent Technologies (2 mentions)
A0082, by Agilent Technologies (2 mentions)
Axio scope a1, by Zeiss (2 mentions)
Observer z1, by Zeiss (2 mentions)
Axioplan 2 microscope, by Zeiss (2 mentions)
Axiocam hrc camera, by Zeiss (2 mentions)
Trueblack, by Biotium (2 mentions)
Lsm 800, by Zeiss (2 mentions)
Prism 5, by GraphPad (2 mentions)
Matlab r2014a, by MathWorks (2 mentions)
Orca flash4.0 lt, by Hamamatsu Photonics (2 mentions)
Axio scan z1, by Zeiss (2 mentions)
Masson s trichrome, by Polysciences (2 mentions)
Axio scan z1 slide scanner, by Zeiss (2 mentions)
Lsm 700, by Zeiss (2 mentions)
Lsm 700 confocal microscope, by Zeiss (2 mentions)
Lsm 880 confocal microscope, by Zeiss (2 mentions)
Ab111101, by Abcam (1 mentions)
39 protocols using zen 2 blue edition
1
Intravital Imaging of Lymph Node B Cell Recruitment
2
Characterization of Platelet-Like Particles
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
In this manuscript, the term (input) cells is used for particles with a diameter ≥ 7 μm counted in a Malassez haemocytometer. Particles ≤ 7 μm are called platelet‐like particles (PLP). Size of particles was measured using the analytical software tools for microscopy (Zen 2, Blue edition, Carl Zeiss). The same term (PLP) is also used for flow cytometry events appearing inside a polygon gate set as a contour around bPLT as source material 23 (Figs S1 a and b). This polygon gate is further referred to as the morphology gate. Particles within the morphology gate that stain positive for both surface receptor CD42b and CD41 (see below) were then defined as platelets (Fig S1 c).
3
Immunohistochemical Analysis of Lung and Liver
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Formalin-fixed paraffin-embedded lung sections were stained with Alcian blue Elastin Van Gieson and immunostained for αSMA (α-smooth muscle actin; 1:150; Dako, M0851), von Willebrand factor (1:300; Dako, A0082), NF-κB (nuclear factor kappa-B; 1:400; Cell Signaling, D14E12), F4/80 (1:100; Abcam, ab111101), iNOS (inducible NO synthase; 1:100; Abcam, ab15323), and CD206 (1:100; R&D Systems, AF2535). As isotype control, IgG and IgG2a were used. Formalin-fixed paraffin-embedded liver sections were stained with anti-F4/80 to confirm macrophage ablation. For immunohistochemical and immunofluorescent staining, a standard protocol was followed. Histological images were visualized using a Zeiss multislide scanning microscope (Imager.Z2; Carl Zeiss, Ltd) with an Axiocam 506 color camera (Zeiss) for immunohistochemical images and MRm camera (Zeiss) for immunofluorescent images. Slides were scanned sequentially using ×20 magnification objective lens, and the analysis was performed in Zen2 blue edition (Zeiss). For all tissue macrophage quantification, positively stained cells were counted in 6 fields of view at ×200 magnification and normalized to the control group.
4
Intravital Imaging of Leukocyte Trafficking
5
Microscopy Monitoring of Cell Differentiation
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Cells after isolation, during differentiation studies and after staining were monitored on inverted light microscope (Observer Z1, Carl Zeiss, Oberkochen, Germany), under phase contrast and bright field. The images were acquired using the camera AxioCam HR (Carl Zeiss, Germany) and the software ZEN 2 blue edition (Carl Zeiss, Germany).
6
Pollen Morphology Characterization by SEM
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Size and shape of four selected pollen species, Iva xanthiifolia, Populus nigra italic, Populus tremuloides, and Populus deltoids (all from: Sigma-Aldrich, St. Louis, MO, USA) were examined through an electron microscope, the EVO HD 15 (Zeiss, Jena, Germany). Samples were sputtered with a thin gold layer using a Q150R ES sputter coater (Quorum technologies, Houston, TX, USA) before imaging at 5 kV and 7.5 mm working distance. A LSM 710 Axio Observer (Zeiss, Jena, Germany) visualized autofluorescence at a wavelength of 488 nm using an EC-Plan-Neofluar 10×/0.30M27 objective (Zeiss, Jena, Germany). ZEN 2 blue edition (Zeiss, Jena, Germany) was used as computer software. Additional wavelengths (405 & 561 nm) were applied when examining the ointment samples. For SEM imaging, MucilAir™ cell inserts were dehydrated with an ethanol (VWR Radnor, PA, USA) series at room temperature (RT) (70-80-90-96 (two times) and 100% for 10 min each), followed by sputter coating.
7
Microscopic Imaging with Apotome 2
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The Ludin chamber loaded with the sample was mounted on an Axio observer Z1 inverted microscope (Zeiss, Germany) equipped with an Apotome 2 module. The image acquisition was performed using a plan-apochromat 63x/1.4 NA oil immersion objective and excitation/emission filters of GFP and DsRed (Zeiss, Germany). Zen 2 blue edition was used for image acquisition (Zeiss, Germany) software. The images were obtained using an Axiocam 506 mono 6 Mp CCD camera with a frame size of 2752p x 2208p (as czi or tif files) at 8-bit depth. The image processing was done using Fiji (NIH, Bethesda, USA) and MetaMorph (Molecular Devices, San Jose, USA).
8
Histopathologic Evaluation of Mouse Tissues
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Mouse livers and lungs were kept in formalin for 24 h. In the following days, the tissues were dissected, placed in tissue embedding cassettes, and submerged in 70% ethanol for dehydration. Tissues were paraffin‐embedded, sectioned, mounted on slides, routinely stained with hematoxylin and eosin, and cover‐slipped allowing histopathologic evaluation by the specialist at IDEXX BioAnalytics. Microscopic changes were graded as to severity utilizing a standard grading system whereby 0 = no significant change, 1 = minimal, 2 = mild, 3 = moderate, and 4 = severe. International Harmonization of Nomenclature and Diagnostic (INHAND) Criteria standards are used for evaluation. The use of numerical grades allows a mechanism to calculate a total score lesion score which can be used to assess the prevalence and severity of tissue changes within and between groups. The slides were imaged using Zeiss Axio Scan.Z1 slide scanner (Zeiss), and the images were cropped using the manufacturer's software (Zen 2 blue edition, Zeiss).
9
Macrophage Response to Biomaterial Particles
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
To study the macrophage response to the materials, RAW 264.7 cells were seeded at a density of 2 × 105 cells per well (in 2 mL of complete DMEM) in a sterile 12-well culture plate (Greiner Bio-One, Kremsmünster Austria). After 24 h of cultivation under standard cell culture conditions, the medium was removed, and the suspension of SC or LC material particles, made with complete DMEM, with and without the addition of 1 ng/mL of LPS, was added to the cells. Two concentrations of materials’ suspensions, 100 and 20 µg/mL, each in three replicates, were tested. Phagocytosis of the material particles was observed after 24 h incubation with material suspensions on an inverted light microscope Axio Observer Z1 (Carl Zeiss, Oberkochen, Germany) under phase contrast. The images were acquired using the camera AxioCam HR in the software ZEN 2 blue edition (Carl Zeiss, Oberkochen, Germany). The macrophage inflammatory-related secretion products were measured in media 24 h after exposure to the materials.
10
Quantification of Hippocampal Immune Cells
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!