Leica dfc495

A cut piece of U. gibba stolon approximately 3 cm long from in vitro culture was placed in liquid 1B medium in a small Petri dish (Sterlin, 50-mm diameter, 124; Slaughter). The youngest trap after emergence from the circinate apex (approximately 150 μm) was imaged every 24 hours until maturity under bright-field light on a Leica M205C stereomicroscope with a Leica DFC495 camera (Leica, Milton Keynes, UK). Trap length (Fig 1A) was measured (Leica LAS version, 4.2 software) and natural log of length plotted against time (Microsoft Excel with LINEST function, Fig 3J, S3 Data). The growth curve was extrapolated back to when the trap was 10 μm long, corresponding to 1–2 cells, which we took to be the initiation stage.

During three field expeditions in September 2007, September and November 2017, a total of fifteen flowering and five fruiting individuals from the type locality were collected from Tianmenshan National Forest Park, Zhangjiajie City, Hunan Province, Southwest China. The information and measurements of the new species were taken from live and dried herbarium specimens and from field data. Seeds were examined and imaged with a Leica M205C stereomicroscope attached to a video camera (Leica DFC495). The morphological comparisons with related species, viz., Swertiabimaculata, S.tashiroi Makino, S.oculata Hemsl., S.tozanensis Hayata, S.cordata (Wall. ex G.Don) C.B.Clarke and S.shintenensis Hayata, are based on herbarium specimens (about 2300 specimens) and relevant literature (Ho et al. 1988 ; Ho and Pringle 1995 ; Rybczynski et al. 2014 (link); Ho and Liu 2015 ). Specimens deposited in the following herbaria were examined: CSFI, CSH, CZH, JIU, HTC, IBK, IBSC, LBG, KUN, PE, SYS and WUK (Thiers 2015 ).
The number of mature individuals was recorded in the field in twenty 1 m² sampling plots. We assessed the preliminary conservation status of the new species using our field knowledge and applying the IUCN (2017) criteria. The taxonomic treatment of the genus Swertia follows Ho and Liu (2015) .

Myocardial fragments were fixed in 4% buffered formalin, processed by a standard technique, and paraffin-embedded sections (3–4 μm thick) were stained with hematoxylin and eosin, Masson’s trichrome (Bio-Optica), Sirius Red (Bio-Optica) and used for histological examination. Masson’s trichrome-stained sections were used in total for morphometric evaluation of percent fibrosis at × 100 using the Image-Pro Plus program (version 6.0.0.260; Media Cybernetics, USA). Sirius Red-stained sections were used for a semi-quantitative 5-point assessment of the amyloid deposits distribution in the interstitium around blood vessels and cardiomyocytes: 0—no deposits; 1—deposits were around single cardiomyocytes; 2—around small groups of cardiomyocytes; 3—around half of cardiomyocytes and single vessels; 4—around more than 75% of cardiomyocytes and blood vessels. The slides were examined by light microscope Leica DMRB with Leica DFC495 camera and Leica PL FLUOTAR 10 × /0.3 and Leica N PLAN 40 × /0.65 (Leica Microsystems Gmbh, Austria).