Cell Image Analysis 10 - · PDF fileDigital Cell Image Analysis Move Away From The Microscope ... Slide Preparation/Staining ... peripheral circulation and to the spleen and other

1

© 2015 Sysmex America, Inc. All rights reserved.

Digital Cell Image Analysis

Move Away From The Microscope

Arby Uy, MT(ASCP)Product Marketing Manager

Sysmex America, Inc.

© 2015 Sysmex America, Inc. All rights reserved.© 2015 Sysmex America, Inc. All rights reserved.

GOALS FOR TODAY

Obtain a working knowledge of digital cell image analysis.

Learn briefly about how automated digital systems work.

Obtain an understanding of some of the advantages of using automated cell image analysis in the clinical laboratory.

Demonstrate how automated cell image analysis can provide better and more rapid diagnoses through the use of examples


BEYOND TECHNOLOGY BEYOND EXPECTATIONS

Quality

Dependability

Accuracy

2


greater accuracy, specificity, productivity

NEXT GENERATION

DIAGNOSTICSsuperior insight and control, enhancing speed to treatment

ADVANCED TOOLS & TECHNOLOGIES

automatically balances work-flow and routine tasks

PROCESS OPTIMIZATION

driving improvement,supporting your staff,

helping you plan for the future

HARMONIZED SUPPORT


LABORATORY TRENDS

Reduction of Qualified Medical TechnologistsMore Generalists

Fewer Specialists

Increased WorkloadsDecreased Staffing


LABORATORY TRENDS

Demand for Accurate Results

Demand for Shorter TAT

Increased Capital Equipment cycles

3


HOW HAS INDUSTRY TYPICALLY ADDRESSED THESE TRENDS ?


OPTIONS TO MANAGE “THE SLIDE”

Review Remains Significantly a Manual Process


CONCERNS WITH MANUAL MICROSCOPY

LIS

Labor intensive

Not standardized

Challenging to train

No historical images

Limited consultation

No traceability

4


WHY AUTOMATE THE MANUAL DIFFERENTIAL

The declining availability of Medical Technologists

The need for a greater level of standardization and consistency for the manual differential

Increasing demand for connectivity between healthcare providers


AUTOMATED CELL IMAGE ANALYSIS

Locates cells on a glass slide

Pre-classify using an artificial neural network

Cells grouped, sorted and displayed for review

Historical archiving of all images

Access to images through lab network


How Cell Image Analyzers Work

5


DETERMINING THE WBC MONOLAYER

Starting at a fixed point in the thick area of the smear.

Moving stepwise towards the thinner part of smear, grabbing 10x images.

Based on the number of RBC contours and the average size of them, the start and end points of the WBC monolayer is determined.

13

Start scanning

Center line

10x images

Image captured at 10x magnification


COLLECTING 100X IMAGES

Using the 100x objective, the system starts grabbing 100x images of the cells in the same order as they were pre-located

End

Start

12 mm

Image captured at 100x magnification


ANALYZING THE IMAGE –ARTIFICIAL NEURAL NETWORK

FEATURES

>350 features :•Form

•Colour

•Texture

•Detection

•Markov

•Wavelets

Se

g

Lym

ph

Mo

no

Eo

Ba

so

Ba

nd

Me

ta

Mye

lo

Pro

mye

lo

Bla

st

NR

BC

Gia

nt

PLT S

mu

dge

Art

efa

ct

Un

ide

nt.

Va

rLy

m

clu

mp

s

6


NEURAL NETWORKS

Logic Based ProgrammingSimilar to Facial Recognition Software

A Neural Network does not learn at the customer site, they are established in a controlled environment.

The Neural Network provides accurate pre-classification of the collected cell images

Powerful tools to improve the Differential Process Cell Identification

Time Savings


EXCELLENT STARTING POINT

Cell Class (n) Correct By DM‐Series Pre‐classification (%)

Segmented neutrophils 3,510 92.5

Band neutrophils 868 57.1

Lymphocytes 2,885 96.4

Monocytes 763 81.4

Eosinophils 231 63.2

Basosphils 50 80.0

Blasts 395 65.1

Immature myeloid cells 627 53.2

Nucleated RBC 165 86.7

Pre-Classification Accuracy

Reference: Kratz, A; Bengsston, H; Casey, J; Keefe, J; Beatrice, G; Grzybek, D; Lewandrowski, K; Van Cott, E : Performance Evaluation of the CellaVision® DM96 System: WBC Differentials by Automated Digital Image Analysis Supported by an Artificial Neural Network. Am J of Clinical Pathology; (2005)124: 770-781


EASY VERIFICATION OF RESULTS

18

7


PRE-CLASSIFIED WBC’S PRESENTED ON SCREEN


ONE CLICK HIGH RESOLUTION DETAIL


LOCATE ABNORMAL CELLS FASTER

8


IMPROVED ACCURACY & PROFICIENCY

View cell classes side by side

Aid cell identification by seeing the “company they keep”

Compare patient cells to custom ref cell library

Using your Patient Population

Your Lab’s staining protocol

Access to historical imagesAbility to track patient changes over time


STANDARDIZED RBC CHARACTERIZATION


STANDARDIZE PLATELET ESTIMATES

9


EXPAND WITH BODY FLUID DIFFERENTIALS


FUTURE ENHANCEMENTS

Optional Software ApplicationAdvanced RBC widens the automation scope of the CellaVision DM-Series analyzers and the Sysmex DI-60 analyzer to deliver a more comprehensive and detailed red blood cell analysis.

Pending FDA Review and Clearance Not available in the United States

Advance RBC Application Software


10


NETWORK CONNECTIVITY





11


BENEFITS OF REAL TIME CONNECTIVITY

Clinicians access to images reduces TAT

Easy saving/sharing of images

Real-time collaboration

Email images anywhere

Access from home

Unlimited image storage

Maintain competency across multiple sites

Centralize expertise

Share staff


ARCHIVING

Want to see cells from last month? Start searching

….or start searching!


DI-60™ Integrated Slide Processing System

12


WHAT IS THE DI-60 ?

New Cell Image Analyzer Result of a collaborative R&D effort between Sysmex Corp and CellaVision

Full Integration for the hematology Work CellCBC

Slide Preparation/Staining

Digital Scan & Pre-classification

Network Compatible with Other DI-60’s or CellaVision DM-Series


superior insight and control, enhancing speed to treatment

ADVANCED TOOLS & TECHNOLOGIES

automatically balances work-flow and routine tasks

PROCESS OPTIMIZATION


SYSMEX DI-60™

13


CELL IMAGE ANALYSIS PORTFOLIO

CellaVision® DM-Series:

A good fit for mid- and high volume labs. Analyzes both blood and body Fluid samples

Sysmex DI-60 Cell Image Analyzer:

Complete integration to the XN-3000 and XN-9000, or in configuration with an SP-10 called the Integrated Slide Processor (ISP).


Case Studies


CASE #1 SYSTEMIC CANDIDA DISEASE

14


SYSTEMIC CANDIDA DISEASE

The Candida yeast is normally harmless and resides in the mouth and the digestive track. When your immune system is lowered or favorable conditions like warm and humid atmosphere are available, Candida grows uncontrollably and becomes an infection.

Systemic Candida is most often caused by a simple infection that goes untreated or insufficiently treated.


CASE #2 PARASITE?


CASE #2 MALARIA ?

15


CASE #2 PLASMODIUM VIVAX


PLASMODIUM VIVAX

Malarial parasites work by digesting red cell proteins and making the RBC membrane less deformable, causing hemolysis, increased splenic clearance, and anemia.

Red cell lysis stimulates release of cytokines and TNF-α. The systemic manifestations of malaria such as headache, fever and rigors, nausea and vomiting, diarrhea, anorexia, tiredness, aching joints and muscles, thrombocytopenia, immunosuppression, coagulopathy, and central nervous system manifestations have been largely attributed to the various cytokines released in response to these parasite and red cell membrane products

P. Vivax makes up 16% of cases reported in US

Not found in West Africa as no Duffy Antigen, which is required for entry in to the RBC.

Characterized by:Low to Normal Platelet Count

Anemia

White blood cell (WBC) counts during malaria are generally characterized as being low to normal, a phenomenon that is widely thought to reflect localization of leukocytes away from the peripheral circulation and to the spleen and other organs, rather than actual depletion or stasis.

In P.Vivax it is common to see more than one stage in the life cycle at the same time in the Peripheral Blood.


CASE # 3 HIGH PERCENTAGE OF BLASTS

16


CASE # 3 PRESENCE OF AUER RODS


ACUTE MYELOID LEUKEMIA (AML)

Auer rods can be seen Classically seen in myeloid blasts of M1, M2, M3, and M4 acute leukemias.

Named for John Auer, an American physiologist (1875-1948).

The increased production of blasts blocks the production of normal marrow cells,

Anemia

Thrombocytopenia

Lack of “normal” white blood cells in the peripheral blood.

AML is the most common acute leukemia that affects primarily adults and accounts for over 1 percent of cancer deaths here in the United States.

AML is initially treated with chemotherapy with the intent of inducing remission.


CASE # 4 ERLICHA (ANAPLASMOSIS)

17


ANAPLASMOSIS

Anaplasmosis is often characterized by sudden high fever, fatigue, muscle aches, headache. The disease can be mild or life-threatening. Severely ill patients can have low white blood cell count, low platelet count, anemia, elevated liver enzymes, kidney failure and respiratory insufficiency. Older people or people with immune suppression are more likely to require hospitalization. Deaths have occurred due to anaplasmosis.

There are two kinds of ehrlichiosis, both of which are caused by tick-borne rickettsial parasites called Ehrlichia that infect different kinds of white blood cells. In HME (human monocytic ehrlichiosis), they infect monocytes. In HGE (human granulocytic ehrlichiosis), they infect granulocytes. HGE was renamed anaplasmosis in 2003. It is likely that the lone star tick transmits HME and that the deer tick transmits HGE.

Ehrlichiosis (HME) was originally thought to be only an animal disease. It was described in humans in 1987 and is now found in 30 states, predominately in the southeast, south-central, and mid-Atlantic states, Europe and Africa.


CASE # 5 WORSTENING MYELOFIBROSIS


CASE # 5 WBC IMAGES -METAMYELOCYTES

18


CASE # 5 WBC IMAGES - MYELOCYTES


CASE # 5 WBC IMAGES - PROMYELOCYTES


MYELOFIBROSIS

Myelofibrosis is currently classified as a myeloproliferative disease Where the replacement of the marrow with collagenous connective tissue fibers occurs.

This replacement impairs the patient's ability to generate new blood cells.

This is a fairly typical progression which will most likely continue with thrombocytopenia, gout, and often an acute leukemia.

Only known cure is a bone marrow transplant. Other treatments are merely supportive.

19


CASE # 6 PARASITE ?


CASE # 6 BABESIA


BABESIOSIS – TICKBORN B. MICROTI

Most cases of babesiosis in the United States occur in the Northeast and upper Midwest

New England, New York State, New Jersey, Wisconsin, and Minnesota.

In the Northeast, babesiosis occurs in both inland and coastal areas, including off-shore islands such as Nantucket and Martha’s Vineyard (Massachusetts); Block Island (Rhode Island); and Shelter Island, Fire Island, and eastern Long Island (New York State).

Other possible ways of becoming infected with Babesiainclude:

Contaminated blood transfusion (no tests have been licensed yet for donor screening)

Transmission from an infected mother to her baby during pregnancy or delivery.

The Centers for Disease Control and Prevention have issued a warning about babesiosis. According to the CDC the illness is transmitted through blood transfusions and has infected at least 122 people since 2000. This was released on Sept. 7, 2011.

20


CASE # 7

• A 47 year old woman who is an avid spelunker came to her doctor with complaints of:

– malaise (a general ill feeling)

– fever

– dry or nonproductive cough

– headache

– shortness of breath

– joint and muscle pains

– chills

• She also noted this sore on her forehead.• The physician ordered a CSF due to

her severe headache.


CASE # 7 (OVERVIEW OF CSF)


CASE # 7 HISTOPLASMOSIS

21


HISTOPLASMOSIS

HistoplasmosisCaused by the fungus Histoplasma capsulatum.

Symptoms of this infection vary greatly, but the disease primarily affects the lungs. Histoplasmosis is common among AIDS patients because of their suppressed immune system.

H. capsulatum grows in soil and material contaminated with bird or bat droppings (guano).

The fungus has been found in poultry house litter, caves, areas harboring bats, and in bird roosts (particularly those of starlings).

Histoplasmosis can be diagnosed by:Samples containing the fungus taken from sputum, blood, or infected organs.

Detection of antigens in blood or urine samples by ELISA or PCR.

A test for antibodies against Histoplasma in the blood.


CASE # 8 SICKLE CELL DISEASE


SICKLE CELL DISEASE

Sickle-cell disease (SCD), or sickle-cell anemiaIs a recessive genetic blood disorder characterized by RBCs that assume an abnormal, rigid, sickle shape.

Sickling decreases the cells' flexibility and results in a risk of various complications. The sickling occurs because of a mutation in the hemoglobin gene.

There are different types of crisis events, including vaso-occlusive, aplastic, and hemolytic.

Crisis events can be caused by physical stress, changes in temperature or seasons, emotional stress, etc. Different people have different triggers. The length of a crisis can vary considerably.

Hemolytic crises characterized by acute, accelerated drops in hemoglobin level. The red blood cells break down at a faster rate. Management is supportive and often involves blood transfusions.

22


CASE STUDIES SUMMARY

Automated Cell Image Analysis in the Clinical Lab can be useful for many reasons including:

Clinical Capabilities

Operational Capabilities

Financial Capabilities


VALUE TO LABORATORY

Improved WorkflowIncreased quality and consistency

Ability to review smears easily and cells classed

Identify issues with technicians and ability to educate

Improved ErgonomicsStaff Comfort

Cell Image adjustments

Competency and EducationStudents

Slide Review

Multiple Databases

Decreases tech time


VALUE TO LABORATORYImproved Accuracy & Proficiency

23


VALUE TO LABORATORY

Cells are automatically located

Pre-classification of cells removes the need to manually search the slide

Display of all WBC on one screen allows fast confirmation of cell counter Diff

Multiple slide merge on low counts eliminates buffy coat preps

Streamlines collaborations & consultations

Eliminates searching for slides

Enhanced Efficiency


VALUE TO LABORATORYDecreased Time for Smear Review

Operator Manual Diff CIA DIFF

A 1:45 1:05

B 1:40 1:10

C 3:45 1:30

D 4:10 1:40

E 3:10 1:14

Conclusions:

“The time saved when using the DM96 is greatest for the less experienced laboratory

scientists. With increased instrument familiarity with the instrument there may be potential for

even more time saved”

Comparison of time taken to complete the 30 differentials on the CellaVision DM96 including reclassification of cells with time taken to perform the same differentials manually

Reference:Briggs C., Longhair I., Slavik M., Thwaite K., Mills R., ThavarajaV., Foster A., Romanin D., Machin S.J. (2009) Can automated blood film analysis replace the manual differential? An evaluation of the CellaVision DM96 automated image analysis system. Jnl. Lab. Hem. 31, 48–60


VALUE TO LABORATORYDecreased Time for Smear Review

Conclusions:

Automated image analysis and classification has now reached a stage where it can meet the requirements of a modern routine hematological laboratory in terms of reliability and efficiency. (The systems) are reliable and accurate and enable automated analysis of blood smears in the hematology laboratory. The systems operate most effectively in screening routine blood samples….

Reference: Ceelie H, Dinkelaar RB, van GW. Examination of peripheral blood films using automated microscopy; evaluation of Diffmaster Octavia and Cellavision DM96. J Clin Pathol 2007 Jan;60(1):72-9.

Figure 4: Results for the manual differential count (n = 220) and the DM96 (n = 315). For each method, the mean time of analysis per sample was reduced by 63% with the automated system.

~ 63% reduction in mean time per smear reviewed

24


VALUE TO PATHOLOGY

Enables Real Time CollaborationRemote Review Software

Pathologist complete reviewOn-site or Off-site

Real time consultations

Email cell images for reviewSpecific cells emailed

Decrease time to search for abnormal samples

Electronic record of abnormalities


IMPACT TO THE INSTITUTION

Expanded Cell Image Analysis Program to Include Their Legacy Campus in Plano, TX

Connected Multiple Cell Imaging Systems Into a Single Production Database

Expanded Laboratory Remote Review Capabilities to the Medical Staff

Medical Service Utilizes the Images for Diagnosis and Patient Follow-up



“This New Infrastructure has allowed the Lab to provide improved services to physicians and patients. Prior to having the connectivity, the laboratory had to prepare and deliver the slide to the physicians. This new system alleviates the need to remember to make extra slides, and deliver them to the physician in a timely manner”…..

“Connectivity has made a huge difference. From a workload standpoint, we have been able to handle a significantly increased workload due to the expansion of the automated cell imaging capability”…..

25



“For the latest month since the implementation of CellaVision at Legacy, our average turn around time was 34.7 minutes on a volume of 739 differentials”…. “Our previous volume before the expansion was ~200/month with a 37.2 minute TAT.”.. “More important the number of sample that fell outside our goal of 1 hour TAT was reduced by more than 50%”

Hung Luu, MD. Department of Pathology

Children’s Medical Center


Questions

Thank You

Documents

Cell Image Analysis 10 - · PDF fileDigital Cell Image Analysis Move Away From The Microscope ... Slide Preparation/Staining ... peripheral circulation and to the spleen and other