Auramine o

One milliliter of the homogenized stool was sedimented, and a smear (1 to 2 cm) was prepared and air dried. The dried smear was stained for 15 min using a 0.5% solution of auramine O (Merck, Darmstadt, Germany), decolorized for 2 min in 3% acid alcohol, and counter stained for 1 min using 0.5% potassium permanganate solution. Smears were examined within 1 h under a fluorescence microscope at ×400 magnification.

The cuticle was stained using Auramine O (Sigma-Aldrich Sp. z o.o., Poznań, Poland), and later, the traps were examined using a Leica DM6000 B (Leica Microsystems GmbH, Wetzlar, Germany) and also a Nikon Eclipse E400 light microscope (Nikon, Tokyo, Japan) with a UV-2A filter (Ex. 330–380 nm, DM. 400 nm, Em. 420-α nm).

Trentepohlia aurea, T. umbrina, and T. jolithus cells were stained with 0.1% Auramine O (Sigma-Aldrich, Steinheim, Germany), 1% Calcofluor white (Sigma-Aldrich, Steinheim, Germany; catalog number: 18909) or Carbotrace 480 (blue), Carbotrace 520 (green) and Carbotrace 540 (yellow) (Ebba Biotech AB, Solna, Sweden) for cell wall staining. Images were taken on an Olympus BX-51 microscope (Olympus, Tokyo, Japan) equipped with an Olympus UC30 digital camera (Olympus, Tokyo, Japan). The microscope is equipped with different filter sets for fluorescence microscopy (WB, WG, WU, and WIB).

Discs (5 mm diameter) were excised from leaves using a cork borer and incubated for 1 h at room temperature with tetramethylrhodamine isothiocyanate (TRITC) : Dextran (0.1 mg ml⁻¹, 150k mw; TDB Consultancy AB, Uppsala, Sweden) and Auramine O (0.01% w/v; Sigma). Images were acquired using a Leica SP8 equipped with white light laser (TRITC: excitation, 561 nm; emission window, 609–631 nm; AuramineO: excitation, 458 nm; emission window, 485–532 nm).

Rhodamine
B (CI 45170) (1),
aniline yellow (CI 11000) (2), Congo red (CI 22120) (3), orange II sodium salt (CI 15510) (4), ponceau
S (CI 27195) (5), xylidine ponceau (CI 16150) (6), auramine O (CI 41000) (7), brilliant green
(CI 42040) (8), malachite green (CI 42000) (9), and naphthol yellow S (CI 10316) (10) were obtained
from Sigma-Aldrich Inc., St. Louis, MO, USA. Methylene blue (CI 52015)
(11), basic fuchsin (CI 42510) (12), methyl
violet 2B (CI 42535) (13), and Martius yellow (CI 10315)
(14) were purchased from Fluorochem Ltd., Hadfield, UK.
Undyed, degummed, unmordanted silk (2-ply, 66 Tex, thread count 43
cm^–2) and undyed, washed, unmordanted wool cloth
(3-ply, 158 Tex, thread count 36 cm^–2) from the
Monitoring of Damage to Historic Tapestries project (MODHT) (FP5,
EC contract number EVK4-CT-2001-00048)^{26 ,27} and undyed
cotton cloth locally purchased (thread count 32 cm^–2) were used as the reference cloths (microscope images of reference
cloths in Figure S1, ESI). The H₂O, MeOH, and CH₃CN (ACN) (LC–MS grade) were purchased
from Fisher Scientific, Waltham, MA, USA. Dye samples from Lehne’s
handbook, 1893 (Figure 2), were also analyzed. Fabric clips (Prym Love clips, 1.0 ×
2.6 cm) were bought locally, while water-sensitive paper (Pentair
Hypro) was purchased from Agratech NW Ltd., Rossendale, UK.

Human type II alveolar pneumocytes A549 (CCL185) and RAW 264.7 cells were separately cultured in 75 cm² culture flasks (CAS, Shanghai, China) with antibiotic-free Dulbecco’s modified Eagle’s medium (HyClone laboratories, Logan, Utah) supplemented with 10% fetal calf serum (Gibco BRL, Grand Island, NY). A549 and RAW 264.7 cells were pre-incubated in the medium for 24 h prior to M. bovis infection. The cells were infected with M. bovis at MOI 10:1 for 24 h. M. bovis was then detected by Auramine O (Sigma) methods according to the manufacturer’s instructions.