Eos 1100d camera

Analyses were carried out on juvenile and mature prothalli cultured on W/S medium without growth regulators. Their stage of maturity was identified by morphological features described by Whittier and Storchova [26 (link)]. For each analysis, at least 10 prothalli were used. Photographs shown in this paper are representative examples (Figure 3). The samples (prothallus) were vacuum-infiltrated with FAE fixative with formaldehyde 37%:100% glacial acetic acid:ethanol 50%, 6.5:3.5:100 mL (v/v/v) for 72 h at room temperature [39 (link)]. Fixed tissue was dehydrated in a series of ethanol concentrations: 40, 50, 70, 90, 100% (v/v) (30 min each bath). After dehydration and wax (Paraplast Plus, McCormic Scientific) embedding 5–10 µm specimens were cut. Sections were stained with PAS (periodic acid-Schiff) or double stained with PAS and NBB (Naphtol Blue Black). PAS specifically stained polysaccharides red, and NBB stained soluble and reserve proteins bluish-black. Observations and slides showing important features were made with a light Zeiss Axio Lab A1 microscope equipped with a Canon EOS 1100D camera. Additionally, some microscopic analyses were based on light blue autofluorescence of mature archegonia and red autofluorescence of chlorophyll with a standard filter set (excitation/emission 358/461 nm) with the same microscope.

The composition of the earwig diet was determined by analysing the gastrointestinal tract contents as described by Kocarek et al. [15 (link)], and the goal was to assess the possibility of predation on termites. The collected insects were immediately stored in ethanol (75%) to stop the digestion of food within the alimentary tract. In the dissection of these specimens, the sternites of the thorax and abdomen were cut using slender forceps, and the oesophagus, crop, proventriculus, and midgut were removed. The gastrointestinal tracts of three specimens of Spirolabia kaja sp. nov. were removed; one was from a male, and two were from females. Permanent microscopic preparations of the alimentary tract contents were made with Hoyer’s solution [16 (link)]. The slides were examined for visually identifiable fragments of food (animal, plant, and fungal), and these were documented using an Olympus CX41 microscope and a Canon EOS 1100D camera. The proportions of different kinds of foods were not determined due to the limited quantity of material and the predominance of indistinguishable small fragments.

AE monitoring of root growth was initially conducted in a glass cell. The glass cell (inner space: 27 cm height, 27 cm wide, 1.2 cm thick; Fig. 4c) was filled with a sandy soil (Winzlerboden), sieved at 2 mm, with a bulk density of 1.4 g.cm⁻³. The Winzlerboden soil was chosen because of its high sand content, to favor the production of AE during the root growth. The glass cell was linked to a hanging water bottle to ensure a constant water content (VWC around 30%). Three maize seeds, germinated prior to the experiment, were planted at the soil surface two days after the beginning of the experiment. The glass cell was fixed on a tilted frame, exploiting root gravitropism in order to better visualize root growth. To monitor the background noise occurring during the experiment, one AE sensor was fixed on a glass plate maintained on the same frame as the glass cell, but not in contact with the soil (sensor S1). Two AE sensors were placed in contact to the back side of the glass cell filled by soil, at a distance of 5 and 20 cm from the top soil surface (sensors S2 and S3). A LED light was used to illuminate the setup and allowed visual monitoring of the root growth by the use of a Canon EOS 1100D camera (Canon Inc., Tokyo, Japan; an image was taken every 30 min).