Mc190 hd camera

Epidermis (Ep), proboscis (Pr), intestine (Int) and oocytes (Oo) were chosen as target organs in the onset of previous works on Eulalia viridis^{21 (link),22 (link)}.These works revealed specialised pigments cells in epidermis and proboscis, as well as scattered pigment granules in epithelial cells of the gut and ooplasm. In the present work, cryopreservation and cryotomy were enforced to evaluate the pigments’ native appearance, i.e., without interference of fixatives and other chemicals used in histological and cytological processing. For the purpose, E. viridis were snap-frozen in liquid nitrogen, sectioned and placed in optimal cutting temperature (OCT) medium in an appropriate tissue mould. The OCT medium containing the tissue was frozen to have a solid support which allowed to cut longitudinal sections with 5–15 μm thick in the cryostat at −20 °C in a cryomacrotome (CM3600 XP, Leica Biosystems). Sections were transferred to pre-adhesivated slides (Thermo Scientific Superfrost Ultra Plus) and stored at −80 °C until observation in a DM 2500 LED model microscope equipped with a MC 190 HD camera (both from Leica). Both brightfield and autofluorescence observations were made. Samples were marked with Hoechst 33258 fluorochrome for DNA counterstaining.

An optical microscope image between crossed polarizers was obtained with a polarizing microscope (Leica DM2700P, Germany) equipped with a Leica MC190 HD camera. 2D WAXD measurements were carried out to monitor the evolution of structures. The X-ray wavelength was 0.154 nm and a Mar345 CCD detector (150 × 150 pixels) was employed to collect the time-resolved 2D WAXD patterns. A bundle of BC–Alg macrofibers were placed in a sample holder perpendicular to the X-ray beam. The distance between the detector (Mar 345) and the sample was 195.00 mm. A typical acquisition time was 60 s. The patterns were corrected for air scattering and background. Fit2D software from the European Synchrotron Radiation Facility was used to analyze the data.

Optical microscopy was performed
using a Leica DME optical microscope mounted with a Leica MC190 HD
camera. Images were acquired by Leica Application Suite 4.12.0 software
with and without polarizers. A drop of either surfactant in oil or
oil foam (∼5 μL) was transferred by a micropipette onto
the middle of a glass slide (76 mm × 26 mm) with a single circular
cavity (15 mm) and then covered gently with a thin glass coverslip.
The glass slide containing the sample was placed in the middle of
a hot stage (Linkam PE120) connected to a digital controller (Linkam
T95PE). All images were analyzed with ImageJ software (1.47 V) calibrated
with a Pyser-SGI Graticule Limited. The number average bubble diameter
of a foam sample was calculated from at least 200 representative bubbles.

The obtained samples were examined by multiple analytic techniques. Optical microscope images were obtained with a polarizing microscope (Leica DM2700P, Germany) equipped with a Leica MC190 HD camera. The morphology of the samples was determined by SEM (Zersss Supra 40) and TEM (JEOL 2010F(s)). The STEM and HRTEM images, and EDX elemental mappings were taken on JEMARM 200 F Atomic Resolution Analytical Microscope with an acceleration voltage of 200 kV. The KPFM characterization was carried out with Atom Force Microscope (Dimension Icon). ICP-AES data was obtained by an Optima 7300 DV instrument. XRD was performed on a Japan Rigaku DMax-γA X-ray diffractometer with Cu Kα radiation (λ = 1.54178 Å). XPS was taken on an X-ray photoelectron spectrometer (ESCALab MKII) with an X-ray source (Mg Kα hυ = 1253.6 eV).

HAE cells that had been differentiated for 7 weeks were fixated in PFA (4%) for 4 h, dehydrated, embedded in paraffin, and sectioned. Six μm sections were deparaffinized and stained according to standard HE procedures. Images were made using the Leica Dmi8 microscope with a 40X objective and Leica MC190HD camera using LAS X 3.4.2 software. Images were cropped and a scalebar was added with Adobe Photoshop version 19.1.19.

For quantification of the number of terminal end buds (TEB) and ductal ends in the mammary glands and the length of the epithelium, #4 mammary glands stained with carmine-alumn whole mount were imaged using a Leica S9i stereomicroscope equipped with an integrated 10 MP camera. The amount of TEBs and ductal ends was quantified from the images based on morphology. The length of the epithelium was measured from the same images using ImageJ with its line tool. The length of the whole epithelial network was recorded.
For quantification of ductal density, images of Hematoxylin and Eosin (H&E)-stained pregnant mammary glands were acquired using a Leica Microscope DM2000 with a Leica MC 190 HD camera with an N Plan 10× objective. The percentage of ductal density was quantified from 10× images by applying a series of Gaussian filtering and thresholding for brightness to separate stained tissue area from the lumen and surrounding adipose tissue using a custom Java script that can be obtained from the authors. The ratio of the red channel to the other channels, balanced with the brightness, was used to further separate Hematoxylin-stained nucleus-rich ductal tissue from Eosin-stained connective tissue, and produce a binary image. A size-based filter was applied to remove single nuclei and debris. The remaining area representing the ducts was calculated for each tissue slide.