Eclipse ni light microscope

A. rhizogenes AR1193 cells carrying the indicated constructs were used to transform L. japonicus Gifu, flot1-1, rinrk1-1, nfr1-1, or nfr5-3 hairy roots on half-strength B5 medium. After 2 weeks, the transformed hairy roots were examined regarding DsRed fluorescence using the LUYOR-3415RG Hand-Held Lamp. The transgenic roots were transferred to a vermiculite and perlite (1:1) mixture and then inoculated with M. loti R7A/lacZ 5–7 days later. The L. japonicus hairy roots transformed with pFlot1:GUS were stained with X-Gluc to visualize gene expression at the indicated time points. For the Flot1 complementation assays, the transgenic roots were stained with X-Gal and the infection events were scored by examining the samples using the Nikon ECLIPSE Ni light microscope at 5 dpi. For the RinRK1, NFR1, or NFR5 complementation assays, the number of nodules was determined at 18 days or 3 weeks after the inoculation with M. loti R7A/lacZ.

Pseudocrangonyx specimens were collected from the groundwater of three Korean caves: Deureune Cave (Fig. 1A), Kwangcheonseon Cave (Fig. 1B), and Hwanseon Cave (Fig. 1C). We fixed and preserved the specimens in 99% ethanol. All specimen appendages were dissected in 80% ethanol and mounted with gum-chloral medium on glass slides under an Olympus SZX7 stereomicroscope (Tokyo, Japan). The specimens were examined using a Nikon Eclipse Ni light microscope (Tokyo, Japan) and were illustrated with the aid of a drawing tube. The body length from the tip of the rostrum to the base of the telson was measured along the dorsal curvature to the nearest 0.1 mm. The nomenclature of the setal patterns on the mandibular palp followed the method described by Stock (1974) . The specimens examined in this study have been deposited in the collection at the Nakdonggang National Institute of Biological Resources, Korea (NNIBR).

Giemsa-stained culture smears were imaged on a DM2000 LED light microscope (Leica) equipped with a DMC2900 camera (Leica). Fluorescence microscopy was performed with an Eclipse Ni light microscope (Nikon) fitted with a C11440 camera (Hamamatsu) or with a D6B fluorescence microscope (Leica) equipped with a DFC9000 GT camera (Leica). For detailed microscopy protocols including quantitative analysis of Fluo-CQ uptake, please refer to SI Appendix.

Three fresh adult specimens of each species were anaesthetized by ether, and their external morphologies were observed under a Olympus SZX16 stereomicroscope. Dorsal and ventral skin samples from the four species of Onchidiidae were dissected into small pieces, fixed in Bouin solution and embedded in paraffin wax [18 (link)]. Sections (5~6 μm) were cut on a Leica RM2035 microtome, stained with hematoxylin-eosin and observed under a Nikon Eclipse Ni light microscope.
Ten sections were selected randomly from each specimen for measurements of the skin, epidermis, dermis, stratum compactum and stratum spongiosum at six sites. The data were analyzed using the software package JMP Version 10.0 (SAS Inc., NC, USA).
For scanning electron microscopy (SEM) analysis of the Onchidiidae species, tissues were fixed in a mixture of methanol and glutaraldehyde for one week and then preserved in 75% alcohol. After this procedure, the materials were washed three times in phosphate buffer (pH 7.0) for 15 min each time, cleaned in an ultrasonic water bath for 2~3 min and then dehydrated in a series of increasingly concentrated ethanol solutions (30%, 50%, 70%, 80%, 90%, and 100% ethanol), with 15 min per solution. Finally, the samples were prepared for SEM using critical-point drying, sputter coated with gold using DMX-220 ion-plating equipment and then examined by SEM.

Fourteen weeks post-CCI, four groups of mice (MMP-9 KO [n = 5], MMP-9 WT [n = 5], MMP-9 OE [n = 5], and MMP-9 WT-OE [n = 5]) were transcardially perfused with chilled (4 °C) PBS, followed by 4% paraformaldehyde in 0.1 M PBS. After staining the coronal sections with Cresyl violet, photographs were taken with a Nikon Eclipse Ni light microscope with a PlanApo 2 × 0.1 objective using ImagePro Plus 7.0 software. The histological lesion area was quantified with ImageJ software and is presented as a ratio between the lesion area and somatosensory cortex. To confirm the functionality of the model, gel zymography was performed in unstimulated (naive) and stimulated (24 h after brain injury) animals to evaluate MMP-9 levels in mice with modified genotypes.

The transgenic P. berghei parasite line pv5-tag-GFP^PV (27 (link)) was imaged live using an AxioImager Z2 epifluorescence microscope equipped with an AxioCam MR3 camera (Zeiss). For mechanical parasite expansion, 1 to 2 µL of infected blood was incubated under a 22 × 40-mm coverslip for several minutes until erythrocyte lysis became apparent. P. falciparum parasites were imaged on an Eclipse Ni light microscope (Nikon) fitted with a C11440 digital camera (Hamamatsu). Immunofluorescence analysis was performed with P. falciparum pv5-3xHA:loxP parasites that were fixed in 4% formaldehyde using rat anti-HA (1:500; Sigma Aldrich) and rabbit anti-SERA5 (1:500) (55 (link)) primary antibodies in combination with appropriate fluorophore-coupled secondary antibodies (1:1,000; Thermo Fisher Scientific). Staining with Lysosensor blue DND-167 (Thermo Fisher Scientific), BODIPY 581/591 C11 (Image-iT Lipid Peroxidation Kit, Thermo Fisher Scientific), and CellROX Green (Thermo Fisher Scientific) was performed according to the manufacturer’s instructions.