The saccharides used in this work (trehalose, lactose, glucose, mannitol, and sucrose) were dissolved in PBS buffer at different concentrations of 2, 4, 6, and 8 g/g of dry lipids. Liposomal suspensions were diluted in an equal volume of each saccharide buffered solution in 50 mL tubes, at 10% of fill volume. As control, liposomal suspension was diluted in equal volume of PBS buffer. A very low freezing temperature seems to avoid damage of nanoparticles and improve the lyoprotective effect, as demonstrated in previous studies using nanoparticles with saccharides (De Jaeghere et al., 2000 (link); Moretton et al., 2012 (link)). Considering these assumptions, all the liposomal suspensions were stored for 6 h at −80°C in a deep freezer and then freeze-dried. When indicated, liposomes were stored in a Corning® CoolCell™, in order to achieve a slow rate of freezing, ~ −1°C/min. The freeze-drying process was performed involved only the primary drying using the Labconco FreeZone 2.5 freeze dryer (Labconco, Kansas City, USA), for 24 h at −50°C in a chamber with 6 Pa (conditions set by the equipment). The reconstitution of freeze-dried liposomes to their original volume was made at room temperature with PBS using a vortex mixer. The samples were equilibrated for 1 h and subjected to further tests.
Freezone 2.5 freeze dryer
Manufactured by Labconco
Sourced in United States
The FreeZone 2.5 Freeze Dryer is a laboratory equipment designed for the freeze-drying process. It has a 2.5 liter capacity and is capable of freeze-drying samples.
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Lab products found in correlation
Krs 5 atr crystal, by PerkinElmer (3 mentions)
Acetonitrile, by Chempur (2 mentions)
Sodium chloride (nacl), by Chempur (2 mentions)
Trisodium citrate, by Chempur (2 mentions)
Sesquihydrate disodium hydrogen citrate, by Chempur (2 mentions)
Anhydrous magnesium sulfate 6, by Chempur (2 mentions)
Anhydrous sodium sulfate 6, by Chempur (2 mentions)
Lts350, by Linkam (1 mentions)
Vcx 500, by Sonics (1 mentions)
Phosphorimager, by Bio-Rad (1 mentions)
1200 series instrument, by Agilent Technologies (1 mentions)
Fluoromax 3 spectrometer, by Horiba (1 mentions)
6230 hr esi tof massspectrometer, by Agilent Technologies (1 mentions)
Gelatin powder, by Nitta Gelatin (1 mentions)
Petroleum ether, by Chempur (1 mentions)
Primary secondary amine, by Agilent Technologies (1 mentions)
Acetonitrile, by Agilent Technologies (1 mentions)
Psa primary secondary amine, by Agilent Technologies (1 mentions)
22 protocols using freezone 2.5 freeze dryer
1
Liposomal Cryoprotection with Saccharides
2
FTIR-ATR Analysis of Sn-STPP Complex
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Example 4
A clear solution comprising 2% stannous fluoride (SnF2) and sodium tripolyphosphate (STPP) with a P:Sn molar ratio of 5 is lyophilized using a Labconco FreeZone 2.5 Freeze Dryer. FTIR-ATR analysis is conducted on freeze dried powder on an extended range Spectrum One Perkin Elmer system featuring a CsI beam splitter, DTGS detector, and single-bounce diamond KRS-5 ATR crystal. Sample is placed directly on the ATR diamond. See
Ethanol is added dropwise to a clear solution comprising 2% stannous fluoride (SnF2) and sodium tripolyphosphate (STPP) with a P:Sn molar ratio of 5 until precipitate forms. Solid is filtered and air dried. FTIR-ATR analysis is conducted on solid on an extended range Spectrum One Perkin Elmer system featuring a CsI beam splitter, DTGS detector, and single-bounce diamond KRS-5 ATR crystal. Sample is placed directly on the ATR diamond. See
FTIR-ATR analysis may be conducted on about 10-100 mg of solid.
3
Purification of Diblock Copolymer
4
Diblock Copolymer Purification Protocol
5
Directional Freezing and Lyophilization for Aerogels
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The respective hydrogels were frozen directionally to ensure pore alignment. This was achieved using a computer-controlled temperature base plate (Linkam LTS350) using cooling with liquid N2 and electrically heated coils to maintain the temperature at −80 °C. The frozen hydrogel were then transferred to a lyophilizer (Labconco FreeZone 2.5 Freeze Dryer) operating at −53 °C and 0.113 mbar, which is below the triple point of water49 . The frozen hydrogels were dried for ~24 h to give the CA aerogel. The label given to the aerogels use an abbreviation according to the immersion solvent (solvent exchange) for the corresponding organogel. For example, an aerogel prepared after immersing organogel in 90 vol.% acetone (AVF = 0.9) is referred to as “0.9 A”.
6
Characterization of Sn-TSPP Complex
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Example 3
A clear solution comprising 2% stannous fluoride (SnF2) and tetrasodium pyrophosphate (TSPP) with a P:Sn molar ratio of 1 is lyophilized using a Labconco FreeZone 2.5 Freeze Dryer. FTIR-ATR analysis is conducted on freeze dried powder on an extended range Spectrum One Perkin Elmer system featuring a CsI beam splitter, DTGS detector, and single-bounce diamond KRS-5 ATR crystal. Sample is placed directly on the ATR diamond. See
Ethanol is added dropwise to a clear solution comprising 2% stannous fluoride (SnF2) and tetrasodium pyrophosphate (TSPP) with a P:Sn molar ratio of 1 until precipitate forms. Solid is filtered and air dried. FTIR-ATR analysis is conducted on solid on an extended range Spectrum One Perkin Elmer system featuring a CsI beam splitter, DTGS detector, and single-bounce diamond KRS-5 ATR crystal. Sample is placed directly on the ATR diamond. See
FTIR-ATR analysis may be conducted on about 10-100 mg of solid.
7
Characterization of Sn-TSPP Complex
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Example 3
A clear solution comprising 2% stannous fluoride (SnF2) and tetrasodium pyrophosphate (TSPP) with a P:Sn molar ratio of 1 is lyophilized using a Labconco FreeZone 2.5 Freeze Dryer. FTIR-ATR analysis is conducted on freeze dried powder on an extended range Spectrum One Perkin Elmer system featuring a CsI beam splitter, DTGS detector, and single-bounce diamond KRS-5 ATR crystal. Sample is placed directly on the ATR diamond. See
Ethanol is added dropwise to a clear solution comprising 2% stannous fluoride (SnF2) and tetrasodium pyrophosphate (TSPP) with a P:Sn molar ratio of 1 until precipitate forms. Solid is filtered and air dried. FTIR-ATR analysis is conducted on solid on an extended range Spectrum One Perkin Elmer system featuring a CsI beam splitter, DTGS detector, and single-bounce diamond KRS-5 ATR crystal. Sample is placed directly on the ATR diamond. See
FTIR-ATR analysis may be conducted on about 10-100 mg of solid.
8
Synthesis of Alginate Nanoparticles via External Gelation
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The alginate nanoparticles (Nano.Alg) were obtained by emulsification using the external gelation method. First, 1.35 g of 1% alginate aqueous solution was mixed with 2.04 g of sunflower oil containing 3% SPAN 80 and homogenized for 1 min using a hand-held homogenizer for emulsification. The emulsion was sonicated for 10 min in pulses (50 s on, 10 s off) in an ice bath using 40% maximum amplitude (VCX500, Sonics, Newtown, CT, USA). While stirring, 1 mL of 0.22 M calcium chloride solution with 0.5% poloxamer 407 was added dropwise. The mixture was sonicated for 5 min in pulses (50 s on, 10 s off), followed by 30 min of magnetic stirring. After these steps, the system was centrifuged for 5 min at 3000 g, the supernatant was removed and isopropanol (P.A.) was added to remove oil residues. Subsequently, 500 μL of 10% trehalose was added prior to freeze-drying for 24 h (FreeZone 2.5 freeze-dryer, Labconco, Kansas, MO, USA) to obtain a homogeneous fine powder of alginate nanoparticles.
MFS-loaded alginate nanoparticles (MFS.Alg) were obtained using the same protocol, except that MFS (3 mg) was added to the alginate aqueous dispersion.24
MFS-loaded alginate nanoparticles (MFS.Alg) were obtained using the same protocol, except that MFS (3 mg) was added to the alginate aqueous dispersion.24
9
Synthesis of PNIPAM Microgels via Emulsion Polymerization
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The synthesis of PNIPAM homopolymer microgels was carried out through the classical and most-often-used method of radical thermo-initiated emulsifier-free precipitation polymerization of NIPAM in water in the presence of the crosslinking agent. The NIPAM monomer concentration in the reaction mixture was 1 wt. %; the cross-linker BIS concentration was 1 mol % in terms of the monomer; the initiator APS concentration was 0.07 wt. %. Polymerization was carried out in a glass reactor in an argon atmosphere at a temperature of 80 °C under continuous stirring at a rate of 600 rpm for 24 h. The aqueous dispersion of the synthesized microgels was gradually cooled to room temperature and purified through dialysis (the dialysis bag pore size was 20 kDa) for two weeks. The purified dispersion was dried via lyophilization using a FreeZone 2.5 freeze dryer (Labconco, Kansas City, MO, USA).
10
Fungal Biomass Harvesting and Drying
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The growth media were separated from the fungal biomass by transferring the fermentation broth with plastic Pasteur pipettes into 15 ml Falcon tubes and the subsequent centrifugation at 13,500 rpm for 15 min at 4 °C. Fungal biomass from Falcon tubes was washed three times with cold distilled water and filtered under vacuum using a Whatman No. I filter paper (GE Whatman, USA). Washed fungal biomass was frozen at − 20 °C and then lyophilized 24 h in a FreeZone 2.5 freeze-dryer (Labconco, USA) at − 50 °C and 0.01 mbar pressure. All samples were stored at − 20 °C until analysis.
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