ASCP MB — Technologist in Molecular Biology

Arrays and Mass Spectrometry

Massively parallel hybridization and mass-based detection: expression, SNP, and chromosomal microarrays (array-CGH and SNP arrays for copy number and loss of heterozygosity), bead arrays, MALDI-TOF genotyping by primer extension mass, and MALDI-TOF identification of microbes by protein fingerprint.

  • 4 min
  • 3 steps
  • 4 questions
  • Lesson 34 of 60

In this lesson

  1. Microarrays
  2. Mass spectrometry
  3. What to take from this
Arrays and Mass Spectrometry

Microarrays

A microarray is massively parallel hybridization: thousands to millions of known probes, each at its own address on a chip 1. A labeled sample is hybridized to the whole array, washed, and scanned; the intensity at each address reports how much of that probe’s target bound 2. Position means identity.

Array Probes detect Uses
Expression transcripts (labeled cDNA/cRNA) expression profiling 3; largely replaced by RNA-seq
SNP / genotyping single-base alleles, by mismatch discrimination or primer extension genome-wide SNPs, pharmacogenomic panels 1
Chromosomal microarray (CMA) copy number across the genome microdeletions and duplications
Methylation bisulfite-converted CpG sites tumor classification

Chromosomal microarray is the first-tier test for unexplained developmental delay, intellectual disability, autism, and multiple congenital anomalies, because it finds submicroscopic deletions and duplications - tens of kilobases rather than the 5-10 megabases a karyotype resolves 1:

  • Array-CGH: patient and reference DNA labeled in two colors and co-hybridized; the ratio at each probe shows gains and losses.
  • SNP arrays: single-sample intensity plus genotype, which reveals loss of heterozygosity, uniparental disomy, and long runs of homozygosity (suggesting consanguinity or a recessive region) - and in tumors, copy-neutral LOH.
  • Blind spots: balanced translocations and inversions (no copy change), low-level mosaicism, and single-base variants.

Examples it detects: 22q11.2 deletion (DiGeorge), 7q11.23 deletion (Williams), 15q11-q13 deletions.

Bead arrays put probes on coded microbeads instead of a flat chip; flow-based readers or high-density bead chips read them. Multiplexed bead assays power some pathogen panels, HLA typing, and large SNP chips.

A microarray grid of spots of varying brightness; a chromosomal microarray log-ratio plot with a run of probes dropping at a 22q11.2 deletion, with resolution of about 5 to 10 Mb for karyotype versus tens of kb for microarray and a list of what arrays miss; and a MALDI-TOF schematic with a mass spectrum, used for single-base-extension genotyping and microbe identification.
Position means identity on an array; mass means identity in MALDI-TOF. Credit: StudyCorner diagram after Buckingham and Green and Sambrook · CC BY 4.0 · Source

Quick check

On a microarray, what tells you which target a spot detects?

Quick check

Why is chromosomal microarray the first-tier test for developmental delay and multiple congenital anomalies?

Quick check

What does a SNP array add beyond array-CGH?

Mass spectrometry

A MALDI-TOF (matrix-assisted laser desorption/ionization time-of-flight) instrument embeds the sample in a matrix, ionizes it with a laser, and measures how long ions take to fly down a tube: lighter ions arrive sooner, giving a mass spectrum 1.

Genotyping by mass: after PCR, a primer ending just before the variant is extended by a single base; the two alleles give products differing by one nucleotide’s mass, which MALDI-TOF separates. Dozens of SNPs can be multiplexed per well, for pharmacogenomics, somatic hotspots, and sample identity panels 1.

Microbe identification: a smear of a colony gives a spectrum dominated by abundant ribosomal proteins - a fingerprint matched against a database to identify the organism in minutes. MALDI-TOF has replaced many biochemical identification panels in clinical microbiology; sequencing (16S, ITS) handles what the database can’t (see Microbial Genetics).

Quick check

How does MALDI-TOF identify a bacterium?

What to take from this

Microarrays read thousands of known probes at fixed addresses. Chromosomal microarray finds microdeletions and duplications far below karyotype resolution - first-tier for developmental delay - and SNP arrays add genotypes for LOH and UPD, but balanced rearrangements stay invisible. MALDI-TOF measures mass: single-base extension products for multiplex genotyping, and protein fingerprints for identifying microbes from a colony in minutes.

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Sources for this lesson
  1. 1
    Lela Buckingham. Molecular Diagnostics: Fundamentals, Methods, and Clinical Applications. 3rd ed. F.A. Davis Company. 2019. verifiedThe standard clinical molecular-diagnostics textbook for MLS/MB programs; author holds MB DLM(ASCP). Covers nucleic-acid chemistry, techniques, lab operations, and applications across infectious disease, oncology, genetics, and identity. Primary topic reference for the ASCP MB program.
  2. 2
    Michael R. Green, Joseph Sambrook. Molecular Cloning: A Laboratory Manual. 4th ed. Cold Spring Harbor Laboratory Press. 2012. verifiedThe classic three-volume molecular-biology methods manual — authoritative for nucleic-acid isolation, electrophoresis, restriction digestion, labeling, and hybridization techniques. Standard-tier topic reference for the techniques courses.
  3. 3
    Bruce Alberts, Rebecca Heald, Alexander Johnson, David Morgan, Martin Raff, Keith Roberts, Peter Walter. Molecular Biology of the Cell. 7th ed. W. W. Norton & Company. 2022. verifiedThe canonical cell/molecular biology textbook; used for nucleic-acid chemistry and the central dogma.