diff --git a/CHANGELOG.md b/CHANGELOG.md
index f4a7b64505..9ca0a01d59 100644
--- a/CHANGELOG.md
+++ b/CHANGELOG.md
@@ -24,6 +24,8 @@ and this project adheres to [Semantic Versioning](https://semver.org/spec/v2.0.0
 
 - `opentelemetry-instrumentation-botocore`: Handle dict input in _decode_tool_use for Bedrock streaming
   ([#3875](https://github.com/open-telemetry/opentelemetry-python-contrib/pull/3875))
+- `opentelemetry-instrumentation-aws-lambda`: Fix ImportError with slash-delimited handler paths
+  ([#3894](https://github.com/open-telemetry/opentelemetry-python-contrib/pull/3894))
 
 ## Version 1.38.0/0.59b0 (2025-10-16)
 
diff --git a/instrumentation/opentelemetry-instrumentation-aws-lambda/src/opentelemetry/instrumentation/aws_lambda/__init__.py b/instrumentation/opentelemetry-instrumentation-aws-lambda/src/opentelemetry/instrumentation/aws_lambda/__init__.py
index ed40d4c1c9..9b450cdf21 100644
--- a/instrumentation/opentelemetry-instrumentation-aws-lambda/src/opentelemetry/instrumentation/aws_lambda/__init__.py
+++ b/instrumentation/opentelemetry-instrumentation-aws-lambda/src/opentelemetry/instrumentation/aws_lambda/__init__.py
@@ -460,6 +460,8 @@ def _instrument(self, **kwargs):
             )
             return
         # pylint: disable=attribute-defined-outside-init
+        # Convert slash-delimited paths to dot-delimited for valid Python imports
+        lambda_handler = lambda_handler.replace("/", ".")
         (
             self._wrapped_module_name,
             self._wrapped_function_name,
diff --git a/instrumentation/opentelemetry-instrumentation-aws-lambda/tests/test_aws_lambda_instrumentation_manual.py b/instrumentation/opentelemetry-instrumentation-aws-lambda/tests/test_aws_lambda_instrumentation_manual.py
index 8fbff76675..a468cb986a 100644
--- a/instrumentation/opentelemetry-instrumentation-aws-lambda/tests/test_aws_lambda_instrumentation_manual.py
+++ b/instrumentation/opentelemetry-instrumentation-aws-lambda/tests/test_aws_lambda_instrumentation_manual.py
@@ -727,6 +727,53 @@ def test_sqs_event_sets_attributes(self):
             MOCK_LAMBDA_CONTEXT_ATTRIBUTES,
         )
 
+    def test_slash_delimited_handler_path(self):
+        """Test that slash-delimited handler paths work correctly.
+
+        AWS Lambda accepts both slash-delimited (python/functions/api.handler)
+        and dot-delimited (python.functions.api.handler) handler paths.
+        This test ensures the instrumentation handles both formats.
+        """
+        # Test slash-delimited format
+        slash_env_patch = mock.patch.dict(
+            "os.environ",
+            {_HANDLER: "tests/mocks/lambda_function.handler"},
+        )
+        slash_env_patch.start()
+        AwsLambdaInstrumentor().instrument()
+
+        mock_execute_lambda()
+
+        spans = self.memory_exporter.get_finished_spans()
+        self.assertEqual(len(spans), 1)
+        self.assertSpanHasAttributes(
+            spans[0],
+            MOCK_LAMBDA_CONTEXT_ATTRIBUTES,
+        )
+
+        slash_env_patch.stop()
+        AwsLambdaInstrumentor().uninstrument()
+        self.memory_exporter.clear()
+
+        # Test dot-delimited format (should still work)
+        dot_env_patch = mock.patch.dict(
+            "os.environ",
+            {_HANDLER: "tests.mocks.lambda_function.handler"},
+        )
+        dot_env_patch.start()
+        AwsLambdaInstrumentor().instrument()
+
+        mock_execute_lambda()
+
+        spans = self.memory_exporter.get_finished_spans()
+        self.assertEqual(len(spans), 1)
+        self.assertSpanHasAttributes(
+            spans[0],
+            MOCK_LAMBDA_CONTEXT_ATTRIBUTES,
+        )
+
+        dot_env_patch.stop()
+
     def test_lambda_handles_handler_exception_with_api_gateway_proxy_event(
         self,
     ):