Leica mz apo

After eDNA sampling but within the same day, traditional aquatic insect collection was conducted using the Surber net survey method. A Surber net of 250-µm mesh size in a 30 × 30-cm quadrat at randomly selected riffle and pool habitats at each site (total collection area: 0.18 m²/reach) was used. Collected invertebrates were placed in 99.5% ethanol and morphologically identified using a stereomicroscope (Leica MZ APO; Leica, Germany) by referring to the identification key for the aquatic insects of Japan (Kawai & Tanida, 2018 ). Because morphological identification was difficult for some aquatic insects, particularly Chironomidae and some Baetidae individuals, population abundance and richness were summarized at the family level.
At the same time as aquatic insect sampling, environmental parameters such as water temperature, electrical conductivity (EC; TOA-DKK CM-21P; Japan), and pH (TOA-DKK HM-20P; Japan) were measured in the field. River water samples of 50 ml were collected from each site to obtain the concentrations of total phosphorus (TP) and total nitrogen (TN) measured in our laboratory using a QuAAtro-2HR (BLTEC Corporation, Japan).

Whole mount in situ hybridization and plastic sectioning were performed as previously described [18 (link)]. The antisense riboprobes were as follows: dct and mitfa of medaka [18 (link)] and sox10 of zebrafish [30 (link)]; and sox10a, sox10b and ltk were synthesized from the plasmid described above using SP6 polymerase (Promega) after restriction enzyme digestion: sox10a and sox10b; EcoRI, ltk; XhoI (NEB). For double fluorescent in situ hybridization, the probes were labeled with digoxigenin or fluorescein, and signals were detected with anti-DIG or anti-Fluor POD-conjugated antibodies with using Tyramide Signal Amplification system (Invitrogen). For sectioning, the stained samples were embedded in Technovit 8100 (Heraeus Kulzer) and sectioned at 10μm thickness. Images of stained embryos were taken using Leica MZ APO with attached AxioCam camera (Zeiss) or Zeiss AXIOImager.M2 with attached Orca-frash 4.0 camera. Images of sections were taken using Zeiss AxioPlan2 with an AxioCam camera. Confocal images were obtained with a Zeiss LSM880 laser-scanning confocal microscope.

The new Rovno amber pleasing fungus beetle specimen was purchased from an amber dealer, who obtained the amber mostly in the Varash district of the Rovno region, the most important source of the new Rovno amber taxa in the last few years [30 (link),32 (link),33 (link),38 (link),40 (link),41 (link),45 (link),50 (link),51 (link),52 (link),56 (link),57 (link),58 (link),59 (link)]. It was included in the clear piece of amber, weighting 7.6 g after the primary treatment. The specimen was cut and polished by Anatoly P. Vlaskin (SIZK) for the best visibility of the fossil to the subrectangular piece 10 × 6.5 × 1.6 mm.
Morphological terminology for the body parts of the beetles is following the work of Leschen [2 ].
Photographs were taken at the Schmalhausen Institute of Zoology, National Academy of Sciences of Ukraine (Kiev, SIZK) using the microscope Leica Z16 APO stereomicroscope equipped with a Leica DFC 450 camera. Dimensions were measured by 10 × 23 Leica 10450023Widefield Adjustable Eyepiece on Leica MZ APO.
Eleven extant specimens cited in the text were collected by late Alexey Yu. Isaev (Ulyanovsk State University, Russia) and are deposited in the Zoological Museum of Lomonosov State University (Moscow, ZMSU).
The holotype is deposited in collection of Schmalhausen Institute of Zoology.