Mass Spectra and Yields of Intact Charged Biomolecules Ejected by Massive Cluster Impact for Bioimaging in a Time-of-Flight Secondary Ion Microscope.

Impacts of massive, highly charged glycerol clusters (≳10(6) Da, ≳ ± 100 charges) have been used to eject intact charged molecules of peptides, lipids, and small proteins from pure solid samples, enabling imaging using these ion species in a time-of-flight secondary ion microscope with few-micrometer spatial resolution. Here, we report mass spectra and useful ion yields (ratio of intact charged molecules detected to molecules sputtered) for several molecular species-two peptides, bradykinin and angiotensin II; two lipids, phosphatidylcholine and sphingomyelin; Irganox 1010 (a detergent); insulin; and rhodamine B-and show that useful ion yields are high enough to enable bioimaging of peptides and lipids in biological samples with few-micrometer resolution and acceptable signals. For example, several hundred molecular ion counts should be detectable from a 3 × 3 μm(2) area of a pure lipid bilayer given appropriate instrumentation or tens of counts from a minor constituent of such a layer.

Multiplexed DNA sequencing-by-synthesis.

We report a new DNA sequencing-by-synthesis method in which the sequences of DNA templates, hybridized to a surface-immobilized array of DNA primers, are determined by sensing the number of nucleotides by which the primers in each array spot are extended in sequential DNA polymerase-catalyzed nucleotide incorporation reactions, each with a single fluorescein-labeled deoxyribonucleoside triphosphate (dNTP) species. The fluorescein label is destroyed after each readout by a photostimulated reaction with diphenyliodonium chloride. A DNA polymerase with enhanced ability to incorporate, and to extend beyond, modified nucleotides is used.

Impact desolvation of electrosprayed microdroplets--a new ionization method for mass spectrometry of large biomolecules.

Impact desolvation of electrosprayed microdroplets (IDEM) is a new method for producing gas-phase ions of large biomolecules. Analytes are dissolved in an electrolyte solution which is electrosprayed in vacuum, producing highly charged micron and sub-micron sized droplets (microdroplets). These microdroplets are accelerated through potential differences approximately 5 - 10 kV to velocities of several km/s and allowed to impact a target surface.

252Cf plasma desorption mass spectrometric study of interactions of steroid glycosides with amino acids.

252Cf Plasma desorption with time-of-flight mass spectrometry (TOF-PDMS) has been applied to comparative studies of the interactions of steroid glycosides (SGs) of the spirostan series with amino acids. SGs can interact with amino acids to form heteroclusters of the type [SG + aminoacid + H](+) and [SG + aminoacid + K](+). It is shown how the affinity of SGs for amino acids depends on the structures of both the SG carbohydrate chain and the SG aglycone, and on the nature of the amino acid side chain.