Fluorescein 5 6 isothiocyanate

Konjac glucomannan was purchased from Wuhan Yangyoudao health industry co., LTD company (Hubei, China). Starch from wheat, α‐amylase Type IX‐A from human saliva (activity 1,000–3,000 units/mg protein), pepsin from porcine gastric mucosa (activity 456 U/mg), amyloglucosidase from aspergillus niger (activity 30–60 units/mg protein), amylose from potato, and 3,5‐Dinitrosalicylic acid and fluorescein 5(6)‐isothiocyanate were purchased from Sigma‐Aldrich Co.

Poly(amidoamine) dendrimer (generation 4.0, G₄‐NH₂, water solution, 10% wt/wt) was purchased from Dendritech. tert‐Butyl nitrite (TBN) and fluorescein 5(6)‐isothiocyanate (FITC, ≥90%) was obtained from Sigma‐Aldrich. PVP (average Mw = 40 kDa), N‐acetyl‐d‐penicillamine (NAP), and chlorin e6 (Ce6) were purchased from Aladdin. All other chemicals and solvents were used without further purification unless mentioned. Cell Counting Kit‐8 (CCK‐8) was obtained from Dojindo Molecular Technologies. Polydimethylsiloxane (PDMS) female molds (height: ~1000 μm, base width: ~500 μm, spacing: ~1000 μm, 5 × 5 array) were purchased from the Research Institute of Zhejiang University‐Taizhou. Optimum cutting temperature (OCT) compounds were provided by Sakura Finetek.

Pasteuria ramosa spores were isolated by homogenizing infected Daphnia in ADaM followed by centrifugation at room temperature, 8000 r.p.m. for 5 min. Bacillus thuringiensis spores were thawed and centrifuged at room temperature, 8000 r.p.m. for 5 min. Pasteuria ramosa or B. thuringiensis pellets were suspended in 0.8 ml of 0.1 M sodium bicarbonate (pH 9.1; Sigma, S5761-500G) with 2 mg ml⁻¹ of fluorescein-5(6)-isothiocyanate (Sigma-Aldrich, St Louis, MO). The samples were then incubated in the dark at room temperature, 1600 r.p.m. shaking for 2 h, followed by centrifugation at room temperature, 8000 r.p.m. for 4 min. The pellet was suspended in 0.8 ml sterile water and centrifuged at room temperature, 8000 r.p.m. for 4 min; the supernatant was then removed. This washing step was repeated three times. Spore suspensions were stored in sterile water at 4°C in the dark for further use.

Silymarin powder (SLM, ≥30% silybin (silybin A + silybin B) was obtained from Sigma Aldrich (https://www.sigmaaldrich.com/catalog/DataSheetPage.do?brandKey=SIGMA&symbol=S0292). The other compounds identified were the flavonolignans silychristin, silydianin, isosilybin A and isosilybin B and the flavanonol taxifolin. The extract title was 54.3% according to our previous work [24 (link)]), d-α-Tocopherol polyethylene glycol 1000 succinate (TPGS), silibinin (≥98%, HPLC), fluorescein 5(6)-isothiocyanate (FITC, ≥90%, HPLC), pepsin, bile salts, pancreatic lipase, pancreatin, phosphate buffered saline BioPerformance Certified pH 7.4 (PBS), Tween^®80, lecithin (≥99%, TLC) lyophilized powder, cholesterol BioReagent (≥99%), 1,1-Diphenyl-2-picrylhydrazyl radical (DPPH), human serum albumin (≥96%, HSA) lyophilized powder and all analytical grade and HPLC grade solvents were obtained from Sigma Aldrich (Saint Louis, MO, USA) with the support of Sigma Aldrich Italia (Milan, Italy). Soluplus^® was a gift from BASF (Ludwigshafen, Germany) with the support of BASF Italia, BTC Chemical Distribution Unit (Cesano Maderno, Monza e Brianza, Italy). Distilled water was obtained from a Simplicity^®UV Water Purification System, Merck Millipore (Darmstadt, Germany).

Anhydrous N,N-dimethylformamide (DMF), anhydrous methanol, and glacial acetic acid were obtained from EMD Millipore Chemicals. Ethyl acetate (99.9%, HPLC Plus), ethanol ( ≥ 99.5%, ACS Reagent), isopropyl alcohol ( ≥ 99.5%, BioReagent), tetrahydrofuran (THF, ≥ 99.9%, for high-performance liquid chromatography (HPLC)), nitric acid (70%, ACS Reagent), sulfuric acid (95.0–98.0%, ACS Reagent), hydrochloric acid (37%, ACS Reagent), sodium bicarbonate (BioReagent), sodium sulfate ( ≥ 99.0, ACS Reagent), palladium on carbon (Pd/C, 10 wt.% loading), potassium hydroxide (90%), zirconium chloride (( ≥ 99.5%, trace metal basis), BPDC (97%), dimethyl biphenyl-4,4’-dicarboxylate (99%), fluorescein-5(6)-isothiocyanate ( ≥ 90%), and RITC (mixed isomers) were obtained from Sigma-Aldrich. Ultra-high-purity grade H₂ gas (Praxair, 99.999% purity) was used for the hydrogenation reaction. All starting materials, reagents, and solvents were used without further purification.

Tannic acid, PEG10k, F-68, KO₂, α-amylase, xanthine oxidase, BSA, fluorescein 5(6)-isothiocyanate, pepsin (from porcine gastric mucosa) and pancreatin (from porcine pancreas) were bought from Sigma-Aldrich. DSPE-PEG2k was obtained from Advanced Vehicle Technology Pharmaceutical Co., Ltd (Shanghai, China). Cy5.5-NHS were purchased from (ApexBio Technology, USA). Cytochrome C was obtained from Sangon Biotech (Shanghai, China). Infliximab was purchased from BioChemPartner (Shanghai, China). Dextran sodium sulfate (molecular weight: 36,000-50,000 Da) were purchased from MP Biomedicals Inc. (USA).

DeepTip^TM probes (silicon nitride, nominal resonance frequency 67 kHz–17 kHz; k = 0.48–0.06 N/m) were produced and supplied by the company Bioactive Surfaces S.L. [38 ]. 3-Aminopropyltriethoxysilane, (purity 99%), and AcroSeal^® was obtained from Acros Organics (Thermo Scientific Chemicals, Beerse, Geel, Belgium). Fluorescein 5(6)-isothiocyanate, 30% hydrogen peroxide solution, 50% glutaraldehyde solution in water, Streptavidin from Streptomyces avidinii, albumin biotinamidocaproyl-labeled bovine lyophilized powder, and poly(ethylene glycol)-(N-hydroxysuccinimide 5-pentanoate)-ether 2-(biotinylamino)ethane (MW = 10,000 Da, which corresponds to an approximate length for the spacer of 50 nm) were provided by Sigma-Aldrich (María de Molina, Madrid, Spain). An amount of 32% w/w ammonia solution was obtained from Scharlau (Sentmenat, Barcelona, Spain). Sodium dodecyl sulfate (SDS) was provided by Fisher Scientific (María de Molina, Madrid, Spain).