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. 2010 Mar 24;11(4):1236-52.
doi: 10.3390/ijms11041236.

Whole cell imprinting in sol-gel thin films for bacterial recognition in liquids: macromolecular fingerprinting

Tally Cohen  1 , Jeanna Starosvetsky, Uta Cheruti, Robert Armon
Affiliations

Whole cell imprinting in sol-gel thin films for bacterial recognition in liquids: macromolecular fingerprinting

Tally Cohen et al. Int J Mol Sci. .

Abstract

Thin films of organically modified silica (ORMOSILS) produced by a sol-gel method were imprinted with whole cells of a variety of microorganisms in order to develop an easy and specific probe to concentrate and specifically identify these microorganisms in liquids (e.g., water). Microorganisms with various morphology and outer surface components were imprinted into thin sol-gel films. Adsorption of target microorganism onto imprinted films was facilitated by these macromolecular fingerprints as revealed by various microscopical examinations (SEM, AFM, HSEM and CLSM). The imprinted films showed high selectivity toward each of test microorganisms with high adsorption affinity making them excellent candidates for rapid detection of microorganisms from liquids.

Keywords: bacteria; biosensors; protozoan parasite; sol-gel; sol-gel films; whole cell imprinting.

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Figures

Figure 1.
Figure 1.
Images of imprinted D. radiodurans: entrapped in SG-film (SEM) (A), AFM (B), HSEM (C) and SG-film cavities left after removal of entrapped bacterial cells (SEM) (D).
Figure 2.
Figure 2.
D. radiodurans specific adsorption onto SG imprinted films as detected by acridine orange staining by epifluorescent microscopy (×1,000). (A) D. radiodurans imprinted SG film(Gram staining and AO); (B) imprinted SG film with D. radiodurans exposed to new planktonic suspension of D. radiodurans (AO staining); (C) Control, non-imprinted sol-gel film exposed to same D. radiodurans suspension (AO staining).
Figure 3.
Figure 3.
SEM micrograph of SG film imprinted with D. radiodurans cells and exposed to fresh D. radiodurans suspension for 30 min.
Figure 4.
Figure 4.
Schematic representation of whole cell imprinting of ormosils thin films through a SG procedure.
Figure 5.
Figure 5.
Silver stained SDS-PAGE gel of SG films imprinted with D. radiodurans and non-imprinted. From left to right: first column: molecular markers; third column: imprinted SG with D. radiodurans; fourth column: D. radiodurans whole cell lysate; fifth column: non-imprinted SG film.
Figure 6.
Figure 6.
SEM micrograph of imprinted SG film with E. coli CN13 (A) and (B) after 30 minutes exposure to new E. coli CN13 suspension.
Figure 7.
Figure 7.
SEM micrograph of imprinted SG film with B. subtilis (A) and (B) after 30 minutes exposure to new B. subtilis suspension.
Figure 8.
Figure 8.
SEM micrograph of imprinted SG film with S. natans (A) and (B) after 30 minutes exposure to new S. natans suspension (B).
Figure 9.
Figure 9.
Images of imprinted SG films with Criptosporidium parvum oosysts and adsorption of new oocysts in suspension. (A) SG imprinted film (left-viewed with SEM; right-viewed with CLSM) and (B) SG imprinted film exposed for 30 minutes to new oocysts suspension (left-viewed with SEM; right viewed with an epifluorescent microscope).
Figure 10.
Figure 10.
Sensitivity of imprinted SG films to reinclude D. radiodurans and E. coli cells (separately) in two different volumes (5 and 100 mL).
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