C H A P T E R
IMMULITE® and IMMULITE 1000
7.11 Arthur L. Babson
The IMMULITE® system is manufactured and distributed
by Siemens Healthcare Diagnostics. IMMULITE, a bench-
top instrument, is illustrated in Fig. 1(a). It is a continuous,
random-access, automated immunoassay analyzer designed
around a proprietary assay tube called a test unit that allows
for thorough and efficient washing of an integral antibody- or
antigen-coated bead by rapidly spinning the tube on its verti-
cal axis. All assays use an alkaline phosphatase enzyme label
that is quantified with a sensitive chemiluminescent substrate.
The assay menu includes 105 analytes and more than 5400
instruments were placed in 70 countries around the world.
In 2001 IMMULITE was discontinued with the intro-
duction of IMMULITE 1000 (Fig. 1(b)), basically the
same instrument but with a new casework incorporating
the computer and touch-screen monitor, as well as bulk
fluids with automatic fluid-level indicators resulting in a
consolidated footprint.
FIGURE 1  (a) The IMMULITE system. (b) IMMULITE 1000 (The color version of this figure may be viewed at www.immunoassayhandbook.com).
© 2013 David G. Wild. Published by Elsevier Ltd. All rights reserved.
http://dx.doi.org/10.1016/B978-0-08-097037-0.00044-0
Typical Assay Protocol
The operator loads samples in barcoded sample cups into
the loading chain of the IMMULITE instrument fol-
lowed by up to five barcoded, assay-specific test units in
any order. All subsequent steps are performed automati-
cally by the IMMULITE immunoassay system. The
IMMULITE dispenses the sample, selects and dispenses
the reagent, incubates, separates bound from free, dis-
penses substrate, and reads signal, all automatically as
follows:
ll Sample and alkaline phosphatase-labeled reagent are
added to a test unit.
ll The reaction is incubated for 30 min at 37 °C and
shaken every 10 s.
ll The bead is washed.
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FIGURE 2  The IMMULITE system, schematic.
ll Chemiluminescent substrate is added.
ll The reaction is incubated for 10 min at 37 °C.
ll The luminescence is read.
Time to first result is 42 min and the maximum through-
put is 120 samples per hour. Throughput varies somewhat
with assay mix and is reduced by sequential assays that
require two 30 min incubations.
Product Features
The layout of IMMULITE is illustrated schematically in
Fig. 2. Access to a chain transporting samples and test
units to the various stations of the instrument is continu-
ously available to the operator. Stat samples can be placed
at the head of the queue without interrupting instrument
operation. The pipettor and reagent carousel can be
paused to allow for the addition of reagents at any time
without interrupting those tests in progress. Up to 12 dif-
ferent reagents can be onboard at any time.
Rapid and thorough washing of the coated bead cap-
tured in the test unit is achieved by spinning of the tube on
its longitudinal axis. Excess sample, reagent, and wash
solution are captured in a coaxial waste sump, which is
integrated into the test unit (Fig. 3).
Following washing of the bead, the chemiluminescent
substrate is added and after an additional 10 min incuba-
tion the signal is read by a photon counting PMT.
IMMULITE 1000 uses Microsoft Windows 2000 oper-
ating software for fast system processing, and the
­touch-screen monitor provides easier software access and
fewer keystrokes. Onboard sample dilution is provided for
selected assays, with positive sample identification using
secondary sample tubes. Remote diagnostics connect the
system via modem to DPC for faster technical support.
Ten languages are supported: English, French, Italian,
German, Spanish, Portuguese, Polish, Turkish, Russian,
and Chinese.
Assay Principle
Both competitive assays for haptens and sandwich assays
for large molecules are run with either antibody- or anti-
gen-coated beads. Alkaline phosphatase is the label for all
assays. The patented axial centrifugal separation of bound
and free label provides low background signal.
Calibration
Each analyte has a stored master calibration curve in the
external system computer which is described by a mathe-
matical equation relating photon counts to concentration.
Monthly calibration with two calibrators adjusts the rela-
tionship for the specific instrument and lot of reagents in
use.
Antibodies
The IMMULITE system uses both polyclonal and mono-
clonal antibodies.

FIGURE 3  The IMMULITE assay protocol.
FIGURE 4  Action of alkaline phosphatase in the IMMULITE system.
Separation
Separation is a key feature of the IMMULITE system
(Fig. 3). After incubation the test units are automatically
transferred to the spin-wash station. The tube is spun rap-
idly on its longitudinal axis and the slight draft angle of the
tube allows for instantaneous and complete transfer of
sample and excess reagent from the tube to a coaxial sump
chamber around the top of the test unit. Several 200 µL
volumes of wash water are added to the tube to wash the
bead and the test unit is spun again after each wash leaving
the tube containing the bead totally free of unbound alka-
line phosphatase label.
CHAPTER 7.11  IMMULITE® and IMMULITE 1000 573
Signal Generation
and Detection
After addition of the substrate, the test units enter a sec-
tion of the instrument that excludes ambient light and
allows for a 10 min incubation at 37 °C for development of
the luminescent signal.
The action of bound alkaline phosphatase on adamantyl
dioxetane phosphate creates the unstable adamantyl diox-
etane anion. Breakdown of this unstable anion creates a
prolonged “glow” of light (see Fig. 4).
When each tube reaches the read station in front of the
photomultiplier tube, photon counts are measured for 1 s
through a 2A neutral density filter which attenuates the
signal by a factor of 100. If counts are below a certain level
the attenuator is automatically removed. This automatic
attenuation increases the dynamic range of the photomul-
tiplier tube 100-fold. Counts are measured for 12 consecu-
tive 1 s intervals.
Data Processing
After discarding highest and lowest counts for each tube
the average counts per second are converted to analyte
concentration by means of the stored calibration curves.
The IMMULITE instrument has sufficient storage capac-
ity to save data should any interruption of the communica-
tion with the external computer occur.
574 The Immunoassay Handbook

Interfacing to Laboratory Babson, A.L., Olson, D.R., Palmieri, T., et al: The IMMULITE assay tube: a new
approach to heterogeneous ligand assay. Clin. Chem. 37, 1521–1522 (1991).

Information System Witherspoon, L.R., Babson, A.L. and Olson, D.R. IMMULITE Chemiluminescent
Immunoassay System. In: Immunoassay Automation: An Updated Guide to Systems.
(ed Chan, D.W.) 102–130 (Academic Press, San Diego, 1996).
The IMMULITE software allows for uploading all results
to a laboratory information system.

Further Reading
Babson, A.L. The Cirrus IMMULITE automated immunoassay system. J. Clin.
Immunoassay 14, 83–88 (1991).
Babson, A.L. An automated random-access immunoassay system. Am. Clin. Lab.
Feb., 12 (1992).